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Succinate Dehydrogenase

Succinate dehydrogenase (SDH) is a key enzyme complex in cellular respiration, catalyzing the oxidation of succinate to fumarate in the tricarboxylic acid cycle.
SDH plays a critical role in energy production and is invovled in various metabolic processes.
Researchers studying SDH can leverage the power of PubCompare.ai, an AI-driven platform that helps optimize experiments by easily locating relevant protocols from published literature, preprints, and patents.
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Most cited protocols related to «Succinate Dehydrogenase»

1
Constraint-Based Modeling of E. coli Metabolism
Cited 29 times
The constructed stoichiometric model of E. coli contains all presently known reactions in central carbon metabolism with 98 reactions and 60 metabolites (Supplementary Table I). To apply FBA, the reaction network was automatically translated into a stoichiometric matrix (Schilling and Palsson, 1998 (link)) by means of a parser program implemented in Matlab (MATLAB®, version 7.0.0.19920 (R14), The MathWorks Inc., Natick, MA). Assuming steady-state mass balances, the production and consumption of each of the m intracellular metabolites Mi is balanced to yield
with
S corresponds to the stoichiometric matrix (m × n) and ν (n × 1) to the array of n metabolic fluxes with νilb as lower and νiub as upper bounds, respectively. The above equations represent the conservation law of mass that is fundamental to constraint-based modeling. For all herein presented stoichiometric analyses, maximization of biomass yield is synonymous to the frequently used maximization of growth rate objective (Price et al, 2004 (link)). This is because stoichiometric models are sets of linear balance equations that are inherently dimensionless, hence maximization of the biomass reaction optimizes the amount of product (i.e., the yield) rather than a time-dependent rate of formation. The P-to-O ratio constraint was implemented by omitting the energy-coupling NADH dehydrogenase I (Nuo), cytochrome oxidase bo3 (Cyo) and/or cytochrome oxidase bd (Cyd) components of the respiratory chain. For a ratio of unity, Cyd and Nuo were set equal to zero. Under anaerobic conditions, electron flow is only possible via the NADH oxidases Nuo or NADH dehydrogenase II (Ndh) to fumarate reductase (Frd), hence coupled to succinate fermentation. For nitrate respiration, the terminal oxidase nitrate reductase (Nar) was used instead of Cyd or Cyo (Unden and Bongaerts, 1997 ).
For the genome-scale analysis we used two recently reconstructed models of E. coli metabolism (Edwards and Palsson, 2000b (link); Reed et al, 2003 (link)). In silico growth was simulated on glucose minimal medium for all six environmental conditions. ADP remained unbalanced, since otherwise formation of adenosine would be carbon-limited. For the proton-balanced model of Reed et al (2003) (link), severe alternate optima occurred in central carbon metabolism given an unlimited proton exchange flux between the cell and the medium and a P-to-O ratio of 2, that is the upper bound of the biologically feasible range of P-to-O ratios (Unden and Bongaerts, 1997 ). To prevent the unlimited production of ATP equivalents through the ATPS4r reaction under this condition, all external protons involved in the respiratory chain and the transhydrogenase reaction were balanced (specifically, we balanced the external protons around the reactions ATPS4r, TDH2, CYTBD, CYTBO3, NO3R1, NO3R2, NADH6, NADH7, NADH8). A P-to-O ratio of 2 was implemented by assuming both the transport of four protons through CYTBO3 and NADH6 across the membrane and the diffusion of four protons through ATPS4r for the formation of one ATP equivalent.
Schuetz R., Kuepfer L, & Sauer U. (2007). Systematic evaluation of objective functions for predicting intracellular fluxes in Escherichia coli. Molecular Systems Biology, 3, 119.
Publication 2007
21-hydroxy-9beta,10alpha-pregna-5,7-diene-3-ol-20-one Adenosine Adjustment Disorders Carbon Cell Respiration Cells Diffusion Electrons Escherichia coli Fermentation Genome Glucose Metabolism NADH Dehydrogenase Complex 1 NADH dehydrogenase II NADH oxidase Nitrate Reductase Nitrates Oxidase, Cytochrome-c Oxidases Protons Protoplasm Respiratory Chain Succinate Succinate Dehydrogenase Tissue, Membrane Unden
2
Evaluation of Candidate Reference Genes in Whitefly Transcriptomics
Cited 16 times
Housekeeping genes from a previous B. tabaci transcriptomic study [29] (link) were selected as candidate reference genes including β-actin (Actin), 18S rRNA (18S), heat shock protein (HSP20, HSP40, HSP70, HSP90), γ-tubulin, 60S ribosomal protein L29 (RPL29), succinate dehydrogenase complex subunit A (SDHA), flavoprotein, glyceraldehyde phosphate dehydrogenase (GAPDH), elongation factor 1 alpha (EF-1α), peptidylprolyl isomeraseA (PPIA), NADH dehydrogenase (NADH), Myosin light chain (Myosin L), and adenosine triphosphate enzyme (ATPase). Primer 5.0 (http://www.premierbiosoft.com/) was used to design primers for qRT-PCR analysis. The validity of these candidate reference genes were evaluated under selected biotic and abiotic conditions described in the following sections.
Li R., Xie W., Wang S., Wu Q., Yang N., Yang X., Pan H., Zhou X., Bai L., Xu B., Zhou X, & Zhang Y. (2013). Reference Gene Selection for qRT-PCR Analysis in the Sweetpotato Whitefly, Bemisia tabaci (Hemiptera: Aleyrodidae). PLoS ONE, 8(1), e53006.
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Publication 2013
Actins Adenosine Triphosphate DNAJB1 protein, human EEF1A2 protein, human Enzymes Flavoproteins Gene Expression Profiling Genes Genes, Housekeeping Glyceraldehyde-3-Phosphate Dehydrogenases Heat-Shock Proteins 70 Heat Shock Proteins HSP90 Heat-Shock Proteins Myosin ATPase Myosin Light Chains NADH NADH Cytochrome c Oxidoreductase Oligonucleotide Primers Protein Subunits RNA, Ribosomal, 18S RPL29 protein, human Succinate Dehydrogenase Tubulin
3
Mitochondrial Health Assessment Protocol
Cited 11 times

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Publication 2018
Biological Assay Biological Processes Biological Stress Biopharmaceuticals Blood Platelets Cells Centrifugation Citrate (si)-Synthase Dietary Supplements DNA, Mitochondrial enzyme activity Enzymes Genome Mitochondria Oxidase, Cytochrome-c Real-Time Polymerase Chain Reaction SDHD protein, human Succinate Dehydrogenase
4
Mitochondrial Health Assessment Protocol
Cited 11 times

Protocol full text hidden due to copyright restrictions

Open the protocol to access the free full text link

Publication 2018
Biological Assay Biological Processes Biological Stress Biopharmaceuticals Blood Platelets Cells Centrifugation Citrate (si)-Synthase Dietary Supplements DNA, Mitochondrial enzyme activity Enzymes Genome Mitochondria Oxidase, Cytochrome-c Real-Time Polymerase Chain Reaction SDHD protein, human Succinate Dehydrogenase
5
Quantitative PCR Analysis of Endometrial, Testicular, and Conceptus Transcripts
Cited 8 times
Real-time RT-PCR experiments were carried out under the consideration of the MIQE guidelines [4 (link)]. RNA samples (2 μg/reaction for endometrial and testicular specimens, and 200 ng/reaction for conceptus specimens) were treated with RNase-free DNase I (Ambion, Austin, TX) for 15 min at 37°C, heat denatured (75°C for 10 min), then reverse transcribed using High Capacity cDNA Reverse Transcription Kit and random hexamers (Applied Biosystems, Foster City, CA). cDNA was purified using the QIAquick® PCR Purification Kit (Qiagen, Germantown, MD) and cDNA concentration was determined via spectrophotometry. Purified cDNA (50 ng) was used for each PCR reaction.
For endometrial specimen, the mRNA expression of four putative reference genes, glyceraldehyde 3-phosphate dehydrogenase (GAPDHP), 18S rRNA (18S), beta-2-microglobulin (B2M), and beta actin (ACTB) and one non-reference gene, solute carrier family 36 member 2 (SLC36A2) were measured by real-time RT-PCR. For testicular samples, the mRNA expression of GAPDH, 18S, B2M, ACTB, Succinate dehydrogenase complex (SDHA), and beta glucoronidase (GUSB) as putative internal control genes and aromatase (Cyp19a1) as non-reference transcript were determined. For conceptus tissue, the expression of GAPDH, 18S, B2M, ACTB, SDHA, and tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein (YWHAZ) as reference gene candidates and Cyp19a1as non-reference genes was assessed using quantitative PCR. Primers specific for the selected transcripts were designed using Jellyfish 3.3.1 (Field Scientific LLC, Lewisburg, PA) and are listed in Table 1. Specificity of the primers was confirmed via sequencing of the PCR products to confirm amplification of the intended target sequence. Primer efficiency was assessed using Linreg http://www.gene-quantification.de to ensure all primers resulted in PCR efficiencies of at least 1.9. Real-time PCR was completed using SYBR Green PCR Master Mix (Applied Biosystems) with the following cycling conditions: 95°C for 10 min; 40 cycles of 95°C for 15 sec, 59°C for 1 min; 55 to 95°C for dissociation. Each PCR was performed in triplicate. Specificity of amplification was monitored by including non-reverse transcribed RNA reactions for each sample and by completing a dissociation analysis at the end of each real-time run to verify the amplification of a single product. Cycle threshold (Ct) values were obtained through the auto Ct function.
Klein C., Rutllant J, & Troedsson M.H. (2011). Expression stability of putative reference genes in equine endometrial, testicular, and conceptus tissues. BMC Research Notes, 4, 120.
Full text: Click here
Publication 2011
Actins Aromatase austin beta-Actin BETA MICROGLOBULIN 2 CYP19A1 protein, human Deoxyribonuclease I DNA, Complementary Endometrium Endoribonucleases Family Member GAPDH protein, human Gene Expression Gene Expression Regulation Genes Glyceraldehyde-3-Phosphate Dehydrogenases Oligonucleotide Primers Proteins Real-Time Polymerase Chain Reaction Reverse Transcription RNA, Messenger RNA, Ribosomal, 18S Spectrophotometry Succinate Dehydrogenase SYBR Green I Testis Tissues Tryptophan Tyrosine 3-Monooxygenase

Most recents protocols related to «Succinate Dehydrogenase»

1
Cytotoxicity of Plant Stem Cell Extracts
Not available on PMC !

Example 7

The MTT Cell Proliferation assay determines cell survival following apple stem cell extract treatment. The purpose was to evaluate the potential anti-tumor activity of apple stem cell extracts as well as to evaluate the dose-dependent cell cytotoxicity.

Principle: Treated cells are exposed to 3-(4,5-dimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT). MTT enters living cells and passes into the mitochondria where it is reduced by mitochondrial succinate dehydrogenase to an insoluble, colored (dark purple) formazan product. The cells are then solubilized with DMSO and the released, solubilized formazan is measured spectrophotometrically. The MTT assay measures cell viability based on the generation of reducing equivalents. Reduction of MTT only occurs in metabolically active cells, so the level of activity is a measure of the viability of the cells. The percentage cell viability is calculated against untreated cells.

Method: A549 and NCI-H520 lung cancer cell lines and L132 lung epithelial cell line were used to determine the plant stem cell treatment tumor-specific cytotoxicity. The cell lines were maintained in Minimal Essential Media supplemented with 10% FBS, penicillin (100 U/ml) and streptomycin (100 μg/ml) in a 5% CO2 at 37 Celsius. Cells were seeded at 5×103 cells/well in 96-well plates and incubated for 48 hours. Triplicates of eight concentrations of the apple stem cell extract were added to the media and cells were incubated for 24 hours. This was followed by removal of media and subsequent washing with the phosphate saline solution. Cell proliferation was measured using the MTT Cell Proliferation Kit I (Boehringer Mannheim, Indianapolis, IN) New medium containing 50 μl of MTT solution (5 mg/ml) was added to each well and cultures were incubated a further 4 hours. Following this incubation, DMSO was added and the cell viability was determined by the absorbance at 570 nm by a microplate reader.

In order to determine the effectiveness of apple stem cell extracts as an anti-tumor biological agent, an MTT assay was carried out and IC50 values were calculated. IC50 is the half maximal inhibitory function concentration of a drug or compound required to inhibit a biological process. The measured process is cell death.

Results: ASC-Treated Human Lung Adenocarcinoma Cell Line A549.

TABLE 7
Results of cytotoxicity of apple stem cell extract on lung cancer cell
line A549 as measured by MTT assay (performed in triplicate).
Values of replicates are % of cell death.
Concentration*replicatereplicatereplicateMean of% Live
(μg/ml)123replicatesSDSEMCells
25093.1890.8690.3491.461.510.878.54
10086.8885.1885.6985.920.870.5014.08
5080.5879.4981.0480.370.800.4619.63
2574.2873.8176.3974.831.380.7925.17
12.567.9868.1371.7569.282.131.2330.72
6.2561.6762.4567.1063.742.931.6936.26
3.12555.3756.7762.4558.203.752.1641.80
1.56249.0751.0857.8052.654.572.6447.35
0.78142.7745.4053.1547.115.403.1252.89

Results: ASC-Treated Human Squamous Carcinoma Cell Line NCI-H520.

TABLE 8
Results of cytotoxicity of apple stem cell extract on lung cancer
cell line NCI-H520 measured by MTT assay (performed in triplicate).
Values of replicates are % of cell death.
Concen-%
tration*replicatereplicatereplicateMean ofLive
(μg/ml)123replicatesSDSEMcell
25088.2889.2987.7388.430.790.4611.57
10078.1379.1978.1378.480.610.3521.52
5067.9869.0968.5468.540.560.3231.46
2557.8358.9958.9458.590.660.3841.41
12.547.6848.8949.3448.640.860.5051.36
6.2537.5338.7939.7538.691.110.6461.31
3.12527.3728.6930.1528.741.390.8071.26
1.56217.2218.5920.5618.791.680.9781.21
0.781 7.07 8.4810.96 8.841.971.1491.16

Results: ASC-treated Lung Epithelial Cell Line L132.

TABLE 9
Results of cytotoxicity of apple stem cell extract on
lung epithelial cell line L132 as measured by MTT assay
(performed in triplicate). Values of replicates are % of cell death.
Concen-rep-rep-rep-Mean%
tration*licatelicatelicateofLive
(μg/ml)123replicatesSDSEMcell
25039.5142.5244.0342.022.301.3357.98
10032.9334.4433.6933.690.750.4466.31
5030.6028.9430.5230.020.940.5469.98
2527.9627.8127.1327.630.440.2572.37
12.525.6225.5525.4025.520.120.0774.48
6.2523.1320.8718.6120.872.261.3179.13
3.12513.3411.0811.8312.081.150.6687.92
1.562 6.56 7.31 9.57 7.811.570.9192.19
0.781 8.06 4.30 3.54 5.302.421.4094.70

Summary Results: Cytotoxicity of Apple Stem Cell Extracts.

TABLE 10
IC50 values of the apple stem cell extracts on the on the target
cell lines as determined by MTT assay.
Target Cell
LineIC50
A54912.58
NCI-H52010.21
L132127.46

Apple stem cell extracts killed lung cancer cells lines A549 and NCI-H520 at relatively low doses: IC50s were 12.58 and 10.21 μg/ml respectively as compared to 127.46 μg/ml for the lung epithelial cell line L132. Near complete anti-tumor activity was seen at a dose of 250 μg/ml in both the lung cancer cell lines. This same dose spared more than one half of the L132 cells. See Tables 7-10. The data revealed that apple stem cell extract is cytotoxic to lung cancer cells while sparing lung epithelial cells. FIG. 6 shows a graphical representation of cytotoxicity activity of apple stem cell extracts on lung tumor cell lines A549, NCIH520 and on L132 lung epithelial cell line (marked “Normal”). The γ-axis is the mean % of cells killed by the indicated treatment compared to unexposed cells. The difference in cytotoxicity levels was statistically significant at p≤05.

Example 9

The experiment of Example 7 was repeated substituting other plant materials for ASC. Plant stem cell materials included Dandelion Root Extract (DRE), Aloe Vera Juice (AVJ), Apple Fiber Powder (AFP), Ginkgo Leaf Extract (GLE), Lingonberry Stem Cells (LSC), Orchid Stem Cells (OSC) as described in Examples 1 and 2. The concentrations of plant materials used were nominally 250, 100, 50, 25, 6.25, 3.125, 1.562, and 0.781 μg/mL. These materials were tested only for cells the human lung epithelial cell line L132 (as a proxy for normal epithelial cells) and for cells of the human lung adenocarcinoma cell line A549 (as a proxy for lung cancer cells).

A549 cells lung cancer cell line cytotoxicity results for each of the treatment materials.

DRE-Treated Lung Cancer Cell Line A549 Cells.

TABLE 11
Triplicate results of cell death of DRE-treated
A549 cells measured by MTT assay.
Percentage of live cells calculated as 100% − Mean of triplicates.
Concentration%
(μg/mL)-DRE-Live
treated A549% of cell deathMeanSDSEMcell
25080.4376.4074.8477.232.891.6722.77
10067.6075.2663.7768.885.853.3831.12
5065.3262.9459.9462.732.701.5637.27
2556.8357.9748.1454.315.383.1145.69
6.2555.5949.6949.1751.483.572.0648.52
3.12551.7648.4545.3448.523.211.8551.48
1.56243.6944.0036.0241.244.522.6158.76
0.78137.4726.1919.5727.749.055.2372.26

AVJ-Treated Lung Cancer Cell line A549 Cells.

TABLE 12
Triplicate results of cell death of AVJ-treated
A549 cells measured by MTT assay.
Percentage of live cells calculated as 100% − Mean of triplicates.
Concentration%
(μg/mL)-AVJ-treatedLive
A549% of cell deathMeanSDSEMcell
25076.8178.1675.8876.951.140.6623.05
10076.4075.2673.7175.121.350.7824.88
5065.3266.1559.9463.803.371.9536.20
2550.1048.4556.6351.734.322.5048.27
6.2547.5246.3846.1746.690.720.4253.31
3.12539.8638.6143.7940.752.701.5659.25
1.56232.4019.7730.5427.576.823.9472.43
0.78120.5015.6332.1922.778.514.9277.23

AFP-Treated Lung Cancer Cell line A549 Cells.

TABLE 13
Triplicate results of cell death of AFP-treated
A549 cells measured by MTT assay.
Percentage of live cells calculated as 100% − Mean of triplicates.
Concentration%
(μg/mL)-AFP-treatedLive
A549% of cell deathMeanSDSEMcell
25086.1387.9986.6586.920.960.5613.08
10079.5081.0682.0980.881.300.7519.12
5073.6072.4671.3372.461.140.6627.54
2568.0167.7066.9867.560.530.3132.44
6.2560.8762.1160.7761.250.750.4338.75
3.12549.4851.7650.7250.661.140.6649.34
1.56240.0641.7247.0042.933.622.0957.07
0.78139.2337.7836.8537.961.200.6962.04

GLE-treated Lung Cancer Cell line A549 Cells.

TABLE 14
Triplicate results of cell death of GLE-treated
A549 cells measured by MTT assay.
Percentage of live cells calculated as 100% − Mean of triplicates.
Concentration%
(μg/mL)-GLE-treatedLive
A549% of cell deathMeanSDSEMcell
25088.4291.4990.4490.121.560.909.88
10084.3983.7783.1683.770.610.3516.23
5079.4781.5876.7579.272.421.4020.73
2573.6072.5471.4072.511.100.6327.49
6.2562.8963.6859.9162.161.991.1537.84
3.12550.1854.4751.8452.162.171.2547.84
1.56246.9344.3043.3344.851.861.0755.15
0.78139.5639.3940.9639.970.870.5060.03

LSC-treated lung cancer cell lines A549 cells.

TABLE 15
Triplicate results of cell death of LSC-treated
A549 cells measured by MTT assay.
Percentage of live cells calculated as 100% − Mean of triplicates.
Concentration
(μg/mL)% Live
LSC treated A549% of cell deathMeanSDSEMcell
25077.5478.8578.2078.200.650.3821.80
10077.1476.0476.5976.590.550.3223.41
5066.4268.5266.8267.251.120.6532.75
2559.8067.2264.1663.733.732.1536.27
6.2550.5348.8248.0749.141.260.7350.86
3.12541.1443.6042.7242.491.240.7257.51
1.56239.4739.7440.6139.940.600.3460.06
0.78138.5531.8336.7935.723.482.0164.28

OSC-treated Lung Cancer Cell line A549 Cells.

TABLE 16
Triplicate results of cell death of OSC-treated
A549 cells measured by MTT assay.
Percentage of live cells calculated as 100% − Mean of triplicates.
Concentration
(μg/mL)% Live
OSC-treated A549% of cell deathMeanSDSEMcell
25070.8465.5771.4969.303.251.8730.70
10048.8150.9157.2852.334.412.5547.67
5046.5949.6053.3349.843.381.9550.16
2538.7740.8136.5838.722.111.2261.28
6.2535.7440.7941.0539.193.001.7360.81
3.12534.5533.6837.0235.081.731.0064.92
1.56233.8633.4427.6331.643.482.0168.36
0.78121.3220.0034.8225.388.214.7474.62

L132 cells (“normal” lung epithelial cell line) cytotoxicity results for each of the treatment materials.

DRE-Treated Lung Epithelial Cell Line L132 cells.

TABLE 17
Triplicate results of cell death of DRE-treated
L132 cells measured by MTT assay.
Percentage of live cells calculated as 100% − Mean of triplicates.
Concentration% of %
(μg/mL)cellLive
DRE-treated L132deathMeanSDSEMcell
25086.6686.6186.6686.640.030.0213.36
10076.2977.3976.8476.840.550.3223.16
5065.9268.1767.0167.031.130.6532.97
2555.5458.9557.1957.231.700.9842.77
6.2545.1749.7347.3747.422.281.3252.58
3.12534.8040.5037.5437.612.851.6562.39
1.56224.4231.2827.7227.813.431.9872.19
0.78114.0522.0617.8918.004.012.3182.00

AVJ-Treated Lung Epithelial Cell Line L132 cells.

TABLE 18
Triplicate results of cell death of AVJ-treated
L132 cells measured by MTT assay.
Percentage of live cells calculated as 100% − Mean of triplicates
AFP-treated lung epithelial cell line L132 cells.
Concentration % of %
(μg/mL)cellLive
AVJ-treated L132deathMeanSDSEMcell
25057.0355.9353.6255.531.741.0044.47
10050.9949.7847.0449.272.031.1750.73
5044.9543.6340.4543.012.311.3456.99
2538.9137.4933.8636.752.601.5063.25
6.2532.8831.3427.2830.502.891.6769.50
3.12526.8425.1920.6924.243.181.8475.76
1.56220.8019.0514.1117.983.472.0082.02
0.78114.7612.90 7.5211.733.762.1788.27

AFP-Treated Lung Epithelial Cell Line L132 cells.

TABLE 19
Triplicate results of cell death of AFP-treated
L132 cells measured by MTT assay.
Percentage of live cells calculated as 100% − Mean of triplicates
AFP-treated lung epithelial cell line L132 cells.
Concentration
(μg/mL)% Live
AFP-treated L132% of cell deathMeanSDSEMcell
25056.1555.4357.1956.260.880.5143.74
10049.9548.2447.6448.611.200.6951.39
5043.7441.0538.0940.962.831.6359.04
2537.5433.8628.5433.324.532.6166.68
6.2531.3426.6718.9925.676.243.6074.33
3.12525.1419.489.4418.027.954.5981.98
1.56218.9412.2910.8714.034.312.4985.97
0.78112.73 5.10 6.81 8.214.002.3191.79

GLE-Treated Lung Epithelial Cell Line L132 cells.

TABLE 20
Triplicate results of cell death of GLE-treated
L132 cells measured by MTT assay.
Percentage of live cells calculated as 100% − Mean of triplicates
AFP-treated lung epithelial cell line L132 cells.
Concentration
(μg/mL)% Live
GLE-treated L132% of cell deathMeanSDSEMcell
25084.4283.2083.0883.570.740.4316.43
10080.0579.2978.5979.310.730.4220.69
5072.7571.5974.1072.811.260.7227.19
2580.0581.8679.9980.631.060.6119.37
6.2568.2670.1368.2668.881.080.6231.12
3.12560.6263.0760.6261.441.410.8238.56
1.56248.0748.7748.8348.560.420.2451.44
0.78146.2745.5746.6746.170.560.3253.83

LSC-Treated Lung Epithelial Cell Line L132 cells.

TABLE 21
Triplicate results of cell death of LSC-treated
L132 cells measured by MTT assay.
Percentage of live cells calculated as 100% − Mean of triplicates
AFP-treated lung epithelial cell line L132 cells.
Concentration
(μg/mL)% Live
LSC-treated L132% of cell deathMeanSDSEMcell
25086.4185.8285.7686.000.350.2014.00
10081.2181.2779.9980.820.720.4219.18
5075.9674.7473.5174.741.230.7125.26
2574.7472.7571.4772.991.650.9527.01
6.2570.1368.3268.2668.901.060.6131.10
3.12554.0358.0553.4455.172.511.4544.83
1.56253.9751.9851.9852.641.150.6647.36
0.78146.7945.6244.9245.78 0.940.54 54.22

OSC-Treated Lung Epithelial Cell Line L132 cells.

TABLE 22
Triplicate results of cell death of OSC-treated
L132 cells measured by MTT assay.
Percentage of live cells calculated as 100% − Mean of triplicates
AFP-treated lung epithelial cell line L132 cells.
Concentration %
(μg/mL)Live
OSC-treated L132% of cell deathMeanSDSEMcell
25061.8462.3760.4461.551.000.5738.45
10054.1453.4452.1053.231.040.6046.77
5042.9442.3040.3241.851.370.7958.15
2535.9434.4833.3134.581.320.7665.42
6.2533.9632.6732.0332.890.980.5767.11
3.12527.4826.2026.7226.800.650.3773.20
1.562 9.80 7.29 7.35 8.151.430.8391.85
0.781 7.29 8.98 8.05 8.110.850.4991.89

Calculated values.

TABLE 23
Calculated IC50 doses (ug/mL) and therapeutic ratios
(IC50 for L132 cells/IC50 for A549 cells) for each
treatment material. Values greater than one indicate
that a material would be more selective in killing cancer
cells than normal cells. ASC results imported from
Example 8. These studies indicate that at least
some of the materials may be effective anti-cancer agents.
ASC has outstanding selectivity compared to other materials.
ASCDREAVJAFPGLELSCOSC
A549 12.589.82211.4811.9811.1 13.733.9 
IC50
L132 127.4656.88 62.6682.6577.6369.26715.38
IC50
Ther.10.15.8 5.56.97.0 0.70.5
Ratio

US11872258B2. Oral cavity treatments (2024-01-16). None [US]. Inventors: Maryam Rahimi [US].
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Patent 2024
14-3-3 Proteins 43-63 61-26 A549 Cells Action Potentials Adenocarcinoma of Lung Aloe Aloe vera Antineoplastic Agents Biological Assay Biological Factors Biological Processes Bromides Cardiac Arrest Cell Death Cell Extracts Cell Lines Cell Proliferation Cells Cell Survival Cytotoxin diphenyl DNA Replication Epistropheus Epithelial Cells Fibrosis Formazans Genetic Selection Ginkgo biloba Ginkgo biloba extract Homo sapiens Lingonberry Lung Lung Cancer Lung Neoplasms Malignant Neoplasms Mitochondria Mitochondrial Inheritance Neoplasms Neoplastic Stem Cells Oral Cavity PEG SD-01 Penicillins Pharmaceutical Preparations Phosphates Plant Cells Plant Leaves Plant Roots Plants Powder Psychological Inhibition Saline Solution SD 31 SD 62 SEM-76 Squamous Cell Carcinoma Stem, Plant Stem Cells Streptomycin Succinate Dehydrogenase Sulfoxide, Dimethyl Taraxacum Tetrazolium Salts
2
Macrophage Proteome Analysis of MoxR1 Function
RAW264.7 and TLR4-deficient (3 × 106) macrophage cells were grown in culture plates. After 4 hrs of growth, cells were stimulated with proteins. Post-treatment completion, cells were rinsed with 1× PBS (RT) twice and treated with 2× SDS dye [25 (link)]. The protein was run in varying percentages of SDS-PAGE gel depending on the size of the proteins to be probed. The protein samples were transferred to the blotting membrane (PVDF) for western blot analysis. The membranes were then probed using the primary antibodies against the respective antigens, such as NFKB1, pP-65, cytochrome C (CytC), LC3BII, SQSTM1, beclin1, pMTOR, pULK1, pAKT, pPI3K, HSP60, pcFOS, tcFOS, pJNK1/2, tJNK1/2, tERK1/2, pERK1/2, tP38, pP38, pyruvate dehydrogenase (PD), succinate dehydrogenase (SDHA), CoxIV and Rab7, GAPDH and β-actin antibodies (Cell Signalling). The location of MoxR1 in various purified cellular materials of M. tb was investigated using the antibody against MoxR1 (αMoxR1) (BEI, Resources, NIAID, NIH, USA). The affinity-purified His-tagged MoxR1 protein was confirmed using a monoclonal mouse anti-His antibody (Sigma, USA). For signal generation, secondary antibodies were used as appropriate. Rapamycin and bafilomycin A1 treatment are used for 6 hrs along with MoxR1 treatment to explore its function in autophagy regulation. The protein samples were then prepared by lysis and processed [7 (link),26 (link)]. The western blotting performed to detect the proteins onto the PVDF membrane. 5% BSA in TBST was used for blocking the PVDF membrane in the western blotting of phosphoproteins. The band intensities of proteins were quantified using ImageJ software. GAPDH, β-actin, tFos, tERK1/2, tP38, or tJNK1/2 was used to normalize protein bands as required.
Quadir N., Shariq M., Sheikh J.A., Singh J., Sharma N., Hasnain S.E, & Ehtesham N.Z. (2023). Mycobacterium tuberculosis protein MoxR1 enhances virulence by inhibiting host cell death pathways and disrupting cellular bioenergetics. Virulence, 14(1), 2180230.
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Publication 2023
2-(2-(2-chloro-3-(2-(3,3-dimethyl-5-sulfo-1-(4-sulfo-butyl)-3H-indol-2-yl)-vinyl)-cyclohex-2-enylidene)-ethylidene)-3,3-dimethyl-1-(4-sulfo-butyl)-2,3-dihydro-1H-indole-5-carboxylic acid Actins Antibodies Antigens Autophagy bafilomycin A1 BECN1 protein, human Cells Cytochromes c GAPDH protein, human Immunoglobulins Macrophage Membrane Proteins Mus Oxidoreductase Phosphoproteins polyvinylidene fluoride Proteins Pyruvates SDS-PAGE Sirolimus Succinate Dehydrogenase Tissue, Membrane Western Blot
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Antibody Selection for Subcellular Proteomics
GR-specific affinity-purified polyclonal (GR-H300) or monoclonal (GR-G5) antibodies commercially provided by Santa Cruz Biotechnology (Inc, Europe, Heidelberg, Germany) were used. Mouse monoclonal antibodies against -α-tubulin, -BcL-xLs, -pyruvate dehydrogenase (PDH), -citrate synthase, -glyceraldehyde 3-phosphate dehydrogenase (GAPDH), -peroxisome proliferator-activated receptor alpha (PPARα), -mitochondrial transcription factor A (mtTFA), -malate dehydrogenase 2 (MDH2), -lactate dehydrogenase (LDH), -cytochrome c (cyt c), -cytochrome c oxidase assembly protein COX15 homolog (COX15), -mitochondrial cytochrome c oxidase subunit 5B (COX5B), -cytochrome c oxidase subunit 1 (COXI), -cytochrome c oxidase subunit 2 (COX2), -NADH dehydrogenase [ubiquinone] 1 alpha subcomplex subunit 13 (Grim 19 or NDUFA13), -NAD(P)H: quinone oxidoreductase 1 (NQO1), and -Beclin 1 (BECN1), and rabbit polyclonal antibodies against -α-tubulin, -phosphoenolpyruvate carboxykinase (PEPCK), and -p65 subunit of NF-κB were obtained from Santa Cruz Biotechnology (Inc, Europe, Heidelberg, Germany). In addition, mouse monoclonal antibodies against -β-actin (Sigma Aldrich, St. Louis, MO, USA), -succinate dehydrogenase (SDH) (Invitrogen, Thermo Scientific, GmbH, Frankfurt, Germany), -caspase 9 (Cell Signaling Technology, Leiden, The Netherlands), -cytochrome c oxidase subunit 4 (COXIV) (Abcam, Cambridge, UK), and -mitochondrial cytochrome c oxidase subunit 2 (COXII) (Invitrogen, Thermo Scientific, GmbH, Frankfurt, Germany) were used. Rabbit polyclonal antibodies against -caspase 3, -Bcl-2, -Bax, and -BCL2/Adenovirus E1B 19 kDa protein-interacting protein 3-like (BNIP3L) purchased from Cell Signalling Technology (Leiden, The Netherlands), and anti-P62/Sequestome 1 (SQSTM1) and anti-microtubule-associated proteins 1A/1B light chain 3B (LC3B) antibodies obtained from MBC BioLabs (San Francisco, CA, USA) and Abcam (Cambridge, UK), respectively, were also used. Finally, a goat polyclonal antibody against mitochondrial NADH dehydrogenase [ubiquinone] iron-sulfur protein II (NDUFS2) was purchased from Thermo Fisher scientific (GmbH, Frankfurt, Germany). Details on the source of antibodies and dilutions used are presented in Table 1.
Karra A.G., Tsialtas I., Kalousi F.D., Georgantopoulos A., Sereti E., Dimas K, & Psarra A.M. (2023). Increased Expression of the Mitochondrial Glucocorticoid Receptor Enhances Tumor Aggressiveness in a Mouse Xenograft Model. International Journal of Molecular Sciences, 24(4), 3740.
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Publication 2023
Actins alpha-Tubulin Anti-Antibodies Antibodies ATP-Dependent Phosphoenolpyruvate Carboxykinase BCL2 protein, human BECN1 protein, human BNIP3 protein, human Caspase 3 Caspase 9 Citrate (si)-Synthase Cytochrome c1 Cytochromes c Cytochromes c2 Glyceraldehyde-3-Phosphate Dehydrogenases Glycoprotein Hormones, alpha Subunit Goat Immunoglobulins Iron-Sulfur Proteins Lactate Dehydrogenase Malate Dehydrogenase MAP1LC3B protein, human Mitochondria Monoclonal Antibodies Mus NAD(P)H dehydrogenase (quinone) 1, human NADH NADH Cytochrome c Oxidoreductase Oxidase, Cytochrome-c Oxidases Oxidoreductase PPAR alpha Proteins Protein Subunits PTGS1 protein, human PTGS2 protein, human Pyruvates Rabbits RELA protein, human Succinate Dehydrogenase Technique, Dilution TFAM protein, human ubidecarenone
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MTT Assay for Cell Viability Assessment
The cell viability was assessed using the standard MTT-dye reduction assay as de-scribed previously [66 (link)] with some modifications [67 (link),68 (link)]. The method is based on the bi-otransformation of the yellow tetrazolium salt MTT (3-(4,5-Dimethylthiazol-2-yl)- 2,5-diphenyltetrazolium bromide) to a violet formazan via the mitochondrial succinate dehydrogenase in viable cells. Briefly: the exponentially growing cells were seeded in 96-well flat-bottomed microplates (Corning Costar Flat Bottom Cell Culture Plate, Corning, New York, NY, USA) at a density of 2 × 103 cells per well. After 24 h incubation at 37 °C, the cells from each cell line were treated with the corresponding biologically active substances alone, in concentrations ranging from 16 to 512 µg/mL, or in combination with the chemotherapy drug cisplatin (Sigma-Aldrich) or the hormone therapy drug tamoxifen (Sigma-Aldrich) for 72 h (see Table 2). At least 8 wells were used for each concentration. In some experiments, the cells from the different cell lines were treated with a combination of a constant concentration of the biologically active substances isolated from R. venosa (corresponding to the IC25 values as shown in Table 3) and serial dilutions of cisplatin or tamoxifen ranging from 16 to 512 µM. In other experiments, the chemotherapeutics (cisplatin or tamoxifen) were mixed with the hemolymph fraction 50–100 kDa and were applied in serial dilutions together (see Figure 5). Cisplatin and tamoxifen were dissolved in DMSO. The maximal final concentration of DMSO in the treating solutions was 2.5%. After a 72 h incubation in 5% CO2 at 37 °C, the medium was changed with a phenol-red-free medium, and MTT (Invitrogen) was added in a final concentration of 0.5 mg/mL. The cells were incubated for 2 h in 5% CO2 at 37 °C. Finally, 100µL DMSO per well was added to dissolve the formed formazan crystals. The measurement of the absorbance of the samples was performed on a Varioskan LUX Multimode Microplate Reader (Thermo Fisher Scientific) at 570 nm. GraphPad Prism software v.8 was used for data analysis.
Petrova M., Vlahova Z., Schröder M., Todorova J., Tzintzarov A., Gospodinov A., Velkova L., Kaynarov D., Dolashki A., Dolashka P, & Ugrinova I. (2023). Antitumor Activity of Bioactive Compounds from Rapana venosa against Human Breast Cell Lines. Pharmaceuticals, 16(2), 181.
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Publication 2023
Anabolism Biological Assay Bromides Cell Culture Techniques Cell Lines Cells Cell Survival Cisplatin Hemolymph Hormones Mitochondrial Inheritance MTT tetrazolium Pharmaceutical Preparations Pharmacotherapy prisma Succinate Dehydrogenase Sulfoxide, Dimethyl Tamoxifen Technique, Dilution Therapeutics Viola
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RNA Extraction and qPCR Analysis in Adipose and Brain Tissue
RNA was extracted from visceral adipose tissue or the cerebral cortex by lysis with TRIzol reagent (Life Technologies, Carlsbad, CA, USA, Cat #15596018), according to the manufacturer’s protocol, except for in adipose tissue. For adipose tissue, tissue was lysed using a dounce homogenizer for 20 strokes. The solution was centrifuged for 10 min, and the top layer (fat) was removed prior to the addition of chloroform. After the completion of the TRI-zol protocol, potential genomic DNA contamination was removed from all samples by an RNase-free DNase treatment (Lucigen, Middlesex, UK, Cat #D9905K). Purified RNA (1 μg) was used for reverse transcription using the iScript synthesis system (Bio-Rad, Hercules, CA, USA, Cat #1708891). The resulting cDNA was used for quantitative PCR using a Bio-Rad CFX96 Touch Real-Time PCR Detection System. Standard PCR cycling protocols were used, consisting of 40 cycles, with denaturation at 95 °C and annealing at 60 °C. The amplification efficiency was estimated from the standard curve for each gene. All primers have an efficiency of 88–115%. Relative quantification of mRNA levels was determined by the ΔΔCt method. All groups were compared to the APOE3 control. All experimental primers were compared to the expression of β-actin in the brain and the average expression of succinate dehydrogenase complex, subunit A (SDHA), and hypoxanthine guanine phosphoribosyltransferase (HPRT) in adipose tissue. The expression of control primers showed no significant differences according to the APOE genotype or diet (2-way ANOVA). Primer sequences (Life Technologies) are listed in Table 1.
Christensen A, & Pike C.J. (2023). Effects of APOE Genotype and Western Diet on Metabolic Phenotypes in Female Mice. Metabolites, 13(2), 287.
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Publication 2023
Actins Anabolism Apolipoprotein E3 Apolipoproteins E Brain Cerebrovascular Accident Chloroform Cortex, Cerebral Deoxyribonucleases Diet DNA, Complementary DNA Contamination Endoribonucleases Genes Genome Genotype Hypoxanthine Phosphoribosyltransferase neuro-oncological ventral antigen 2, human Oligonucleotide Primers Protein Subunits Reverse Transcription RNA, Messenger Succinate Dehydrogenase Tissue, Adipose Tissues Touch trizol Visceral Fat

Top products related to «Succinate Dehydrogenase»

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Rneasy mini kit by Qiagen
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The RNeasy Mini Kit is a laboratory equipment designed for the purification of total RNA from a variety of sample types, including animal cells, tissues, and other biological materials. The kit utilizes a silica-based membrane technology to selectively bind and isolate RNA molecules, allowing for efficient extraction and recovery of high-quality RNA.
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TRIzol reagent is a monophasic solution of phenol, guanidine isothiocyanate, and other proprietary components designed for the isolation of total RNA, DNA, and proteins from a variety of biological samples. The reagent maintains the integrity of the RNA while disrupting cells and dissolving cell components.
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Cytochrome c oxidase assay kit by Merck Group
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The Cytochrome c Oxidase Assay Kit is a laboratory tool used to measure the activity of the enzyme cytochrome c oxidase. Cytochrome c oxidase is a key component of the electron transport chain in mitochondria and plays a crucial role in cellular respiration. The assay kit provides a method to quantify the activity of this enzyme in biological samples.
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Mitochondrial complex 3 activity assay kit by Abcam
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The Mitochondrial Complex III Activity Assay Kit is a laboratory tool designed to measure the activity of Complex III, also known as the cytochrome c reductase, within the electron transport chain of mitochondria. This kit provides a quantitative assay to evaluate the enzymatic activity of Complex III in biological samples.
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Ab14744 by Abcam
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Ab14744 is a primary antibody that targets a specific protein. It can be used in various laboratory techniques, such as Western blotting, immunohistochemistry, and enzyme-linked immunosorbent assay (ELISA).
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Succinate dehydrogenase activity colorimetric assay kit by Abcam
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The Succinate Dehydrogenase Activity Colorimetric Assay Kit is a laboratory tool used to measure the activity of the enzyme succinate dehydrogenase. The kit provides a colorimetric-based method for the quantitative determination of succinate dehydrogenase activity.
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Ultrospec 5000 by GE Healthcare
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Rotenone is a naturally occurring insecticide and piscicide derived from the roots of certain tropical plants. It is commonly used as a research tool in laboratory settings to study cellular processes and mitochondrial function. Rotenone acts by inhibiting the electron transport chain in mitochondria, leading to the disruption of cellular respiration and energy production.
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Antimycin A is a chemical compound that acts as a potent inhibitor of mitochondrial respiration. It functions by blocking the electron transport chain, specifically by interfering with the activity of the cytochrome bc1 complex. This disruption in the respiratory process leads to the inhibition of cellular respiration and energy production within cells.
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Citrate synthase assay kit by Merck Group
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The Citrate Synthase Assay Kit is a laboratory instrument designed to measure the activity of the enzyme citrate synthase. Citrate synthase is a key enzyme involved in the citric acid cycle, a metabolic pathway that plays a crucial role in cellular energy production. The assay kit provides the necessary reagents and protocols to quantify citrate synthase activity in various biological samples.

More about "Succinate Dehydrogenase"

Succinate dehydrogenase (SDH), also known as respiratory complex II, is a critical enzyme complex involved in cellular respiration and energy production.
SDH catalyzes the oxidation of succinate to fumarate in the tricarboxylic acid (TCA) cycle, a key metabolic pathway that generates ATP through oxidative phosphorylation.
SDH is composed of four subunits and contains both iron-sulfur clusters and a covalently bound cofactor, flavin adenine dinucleotide (FAD), which are essential for its enzymatic activity.
This complex enzyme plays a pivotal role in mitochondrial function, linking the TCA cycle to the electron transport chain.
Disruptions in SDH activity can have significant impacts on cellular metabolism and energy homeostasis, contributing to various disease states.
Researchers studying SDH can leverage powerful tools like the RNeasy Mini Kit and TRIzol reagent to isolate and analyze RNA from cells or tissues, providing insights into SDH gene expression.
Functional assays such as the Cytochrome c Oxidase Assay Kit and Mitochondrial Complex III Activity Assay Kit can be used to measure SDH activity and assess mitochondrial respiratory function.
Antibodies like Ab14744 can aid in the detection and quantification of SDH subunits.
The Succinate Dehydrogenase Activity Colorimetric Assay Kit provides a convenient way to directly measure SDH activity, while the Ultrospec 5000 spectrophotometer can be used to analyze various mitochondrial and metabolic parameters.
Pharmacological modulators like Rotenone and Antimycin A can be employed to investigate the effects of SDH inhibition on cellular processes.
For a comprehensive understanding of SDH and its role in cellular metabolism, the Citrate Synthase Assay Kit can be used to assess the activity of another key TCA cycle enzyme.
By leveraging these tools and techniques, researchers can gain valuable insights into the regulation and function of SDH, ultimately advancing our understanding of energy production and metabolic homeostasis.