HEK293T cells were lysed 48 h post-transfection with IDH2 WT and mutant constructs under normal and/or hypoxic conditions (200 µM CoCl2 treatment). Changes in intracellular lactic acid and glucose levels were determined with the Lactic Acid Assay Kit (Jiancheng Bioengineering Institute, Nanjing), the Glucose Assay Kit (Rongsheng Biological Pharmaceutical Co. Shanghai), and the ATP Detection Kit (Beyotime Biotechnology Co. Shanghai). According to the manufacturer`s instructions, the changes in the absorption value at 505 nm (glucose) or 530 nm (lactic acid) were measured by UV spectrophotometry (UV-1800PC, MAPADA) after the lysate proteins (5.2 μg/μL) were added to the working solution. ATP levels were measured by Luminometer (Lumat LB 9507, Berthold technologies) after the lysate proteins (2 μg/μL) were added to the working solution. All assays were performed in triplicate. The data were finally analyzed and exported by GraphPad Prism 9.0.0 (GraphPad Software, LLC).
Uv 1800pc
Manufactured by MAPADA
Sourced in China
The UV-1800PC is a UV-Vis spectrophotometer designed for accurate and reliable absorbance measurements. It features a compact, space-saving design and a user-friendly interface. The instrument provides a core function of measuring the absorbance of samples across the ultraviolet and visible light spectrum.
Automatically generated - may contain errors
Lab products found in correlation
Jxfstprp 24, by Jingxin (2 mentions)
Prism 9, by GraphPad (1 mentions)
Atp detection kit, by Beyotime (1 mentions)
Lactic acid assay kit, by Nanjing Jiancheng (1 mentions)
Dr 3900 spectrophotometer, by HACH (1 mentions)
Fe20 plus, by Mettler Toledo (1 mentions)
Toc vcpn, by Shimadzu (1 mentions)
Jem 1400 plus transmission electron microscope, by JEOL (1 mentions)
Malvern zetasizer, by Malvern Panalytical (1 mentions)
Li 6400 portable photosynthesis system, by LI COR (1 mentions)
Le438 ph electrode, by Mettler Toledo (1 mentions)
Tissuelyser 2, by Qiagen (1 mentions)
A045 2, by Nanjing Jiancheng (1 mentions)
Q2000 calorimeter, by TA Instruments (1 mentions)
Id 3003 laboratory x ray generator, by GE Healthcare (1 mentions)
Ir 200, by Thermo Fisher Scientific (1 mentions)
Amino acid analyzer, by Hitachi (1 mentions)
Ezchrom elite software, by Hitachi (1 mentions)
33 protocols using uv 1800pc
1
Intracellular Metabolic Changes in IDH2 Mutant HEK293T Cells
2
Comprehensive Water Quality Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total chlorine and free chlorine were determined immediately after sampling based on the dipropyl-p-phenylenediamine (DPD) method (Spectroquant® chlorine test, EPA 330.5, US Standard Methods 4500-Cl2 G, and EN ISO 7393). The combined chlorine was the subtraction between total chlorine and free chlorine. The conductivity was monitored by a potable conductivity meter (Myron L's Ultrameter II 4P, USA). The solution pH was determined by a pH meter (FE20 Plus, Mettler Toledo). The UV254 was determined by an ultraviolet spectrophotometer (UV-1800PC, Mapada) running under a wavelength of 254 nm. The turbidity was measured by a turbidity meter (HACH 2100Q, UAS). TOC and DOC were determined by a TOC analyser (TOC-V CPN, Shimadzu, Japan). TN was determined by a DBR200 digester and a DR3900 spectrophotometer (HACH, USA).
3
DPPH Radical Scavenging Activity
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity was evaluated according to the protocol of Umayaparvathi et al. (20 (link)). Concisely, 2 mL of protein hydrolysate sample (2, 4, 6, 8, 10 and 14 mg/mL) were added to 2 mL of 0.16 mM DPPH methanolic solution. The mixture was vortexed for 1 min and left to stand at room temperature for 30 min in a dark place, and the absorbance was read at 517 nm (UV-1800PC; Shanghai Mapada Instruments Co., Ltd, Shanghai, PR China). The ability to scavenge the DPPH radical was calculated using the following equation:
where A1 is the absorbance of the control (DPPH solution), and A2 is the absorbance of the sample (DPPH solution with sample). BHT was used as a positive control.
where A1 is the absorbance of the control (DPPH solution), and A2 is the absorbance of the sample (DPPH solution with sample). BHT was used as a positive control.
4
ABTS Radical Scavenging Assay of Protein Hydrolysates
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) free radical scavenging assay was analysed using the method of Chi et al. (12 (link)) with slight modifications. Briefly, ABTS free radical was generated by mixing a concentration of ABTS stock solution (0.007 M potassium persulphate and 0.00245 M ABTS). The mixture was kept in the dark at room temperature for 16 h. The ABTS radical stock solution was diluted in 0.005 M phosphate-buffered saline (PBS) at pH=7.4 to an absorbance of 0.70±0.02 at 734 nm. A volume of 4 mL of diluted ABTS•+ solution was mixed with 0.1 mL of different concentrations of protein hydrolysates (0.5, 1, 1.5 and 2 mg/mL), and incubated at 25 °C for 10 min in a dark place. The absorbance of the mixture was measured at 734 nm (UV-1800PC; Shanghai Mapada Instruments Co., Ltd), and BHT was used as the positive control. The ABTS activity was calculated using the following equation:
where A1 is the absorbance of control and A2 the absorbance of the sample.
where A1 is the absorbance of control and A2 the absorbance of the sample.
5
Ferrous Ion Chelating Assay of Hydrolysates
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The Fe2+ chelating ability of the hydrolysates was evaluated by the method described by Naqash and Nazeer (4 (link)) with minor modifications. Concisely, 3.2 mL of each sample were prepared at the concentrations of 1, 5, 10, 15 and 20 mg/mL and mixed with 40 µL of 0.002 M FeCl2, then the mixture was vortexed for 1 min. A volume of 80 µL of 5 mM 3-(2-pyridyl)-5,6-bis(4-phenyl-sulfonic acid)-1,2,4-triazine (ferrozine) was added to the mixture and then it was incubated at 25 °C for 15 min in a dark place. After incubation, the absorbance at 562 nm was measured with spectrophotometer (UV-1800PC; Shanghai Mapada Instruments Co., Ltd). BHT was used as a positive control. Chelating activity (%) was calculated by the following equation:
where Acontrol is the absorbance of the control and Asample is the absorbance of the sample.
The IC50 value of the hydrolysates for antioxidant parameters such as DPPH, ABTS, •OH, Fe2+ and BHT was determined by linear regression analysis (standard calibration curve) from a plot of concentration against the percentage of inhibition.
where Acontrol is the absorbance of the control and Asample is the absorbance of the sample.
The IC50 value of the hydrolysates for antioxidant parameters such as DPPH, ABTS, •OH, Fe2+ and BHT was determined by linear regression analysis (standard calibration curve) from a plot of concentration against the percentage of inhibition.
6
FRAP Assay for Antioxidant Potential
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The ferric reducing antioxidant power (FRAP) was analysed according to the procedure by Umayaparvathi et al. (20 (link)) with slight modifications. Concisely, 2 mL of protein hydrolysates were taken at various concentrations (0.5, 1, 1.5, 2, 2.5 and 3 mg/mL) and mixed with 2 mL of phosphate buffer (200 mM, pH=6.6), and 2 mL of 1% potassium hexacyanoferrate were added. The mixture was mixed vigorously by vortex mixer (XW-80A; Ningbo Hinotek Technology Co., Ltd., Zhejiang, PR China) for 1 min and incubated at 50 °C for 25 min. Then, 1 mL of 10% trichloroacetic acid was added and mixed, then centrifuged (microcentrifuge D3024R; Scilogex, Beijing, PR China) at 10 000×g for 15 min. After that, the upper layer of the solution (supernatant, 2 mL) was collected and mixed with 2 mL of ultrapure water, and 0.4 mL of 0.1% FeCl3 was added. Finally, the mixture was incubated for 10 min at 25 °C and absorbance was measured by spectrophotometer (UV-1800PC; Shanghai Mapada Instruments Co., Ltd) at 700 nm. BHT was used as a positive control.
7
Enzymatic Activity of Recombinant Protein
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To explore whether the recombinant protein has β-1,3-glucanase activity. The enzyme activity of the recombinant protein was determined by measuring the hydrolysis of laminarin as a substrate. The reaction mixture containing 1% laminarin and 1 nmol of (0.2 mg/mL) recombinant protein was incubated in 50 mM sodium acetate buffer (pH 6.0) for 30 min at 40 • C. Subsequently, 2 mL of 3,5-dinitrosalicylic acid (DNS) solution was added at 100 • C for 10 min and finally placed on ice for 2 min to terminate the reaction. The released amount of reducing sugar was measured using a spectrophotometer (UV-1800PC, Mapada, Shanghai, China) at 520 nm. One unit (U) of enzyme activity was defined as the amount of enzyme that released 1 µmol of glucose per minute under the above-measured conditions.
To evaluate the optimal pH of the enzyme activity of the recombinant protein, the enzyme activity of the recombinant protein was measured in two buffers (50 mM) with different pH: sodium acetate (pH 3, 4, 5, and 6) and Tris-HCl (pH 7, 8, and 9). The effect of temperature on the enzyme activity of the recombinant protein was measured at different temperatures (30, (link)40, (link)50, 60 , 70 and 80 • C) in 50 mM sodium acetate buffer (pH6.0). The enzyme activity assay was completed with three replicates.
To evaluate the optimal pH of the enzyme activity of the recombinant protein, the enzyme activity of the recombinant protein was measured in two buffers (50 mM) with different pH: sodium acetate (pH 3, 4, 5, and 6) and Tris-HCl (pH 7, 8, and 9). The effect of temperature on the enzyme activity of the recombinant protein was measured at different temperatures (30, (link)40, (link)50, 60 , 70 and 80 • C) in 50 mM sodium acetate buffer (pH6.0). The enzyme activity assay was completed with three replicates.
8
Photocatalytic Degradation of Rhodamine B
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The photocatalytic activity of the catalysts as prepared was evaluated through experiments involving the photocatalytic degradation of RhB under visible light exposure. The light source used was a 300 W Xe lamp with a cut-off filter (λ > 420 nm). To reach an adsorption–desorption balance, photocatalysts (100 mg) were distributed in 200 mL of RhB (20 mg L−1) reaction solution, which was stirred in a lightless environment for 30 minutes. At regular intervals, 3 mL of the suspension was collected and centrifuged (8000 rpm, 10 min) to separate the catalysts for further analysis. A UV-vis spectrophotometer was used to measure the concentration of Rhodamine B (MAPADA, UV-1800PC).
To assess the photocatalytic stability of the AT2 nanocomposite, cycle photocatalytic studies were carried out using the aforementioned procedures. The catalyst was gathered after every cycle of the photocatalytic procedure, cleaned with deionized water, and then re-dispersed into a new RhB solution for the following cycle. Trapping agent studies were used to investigate the photocatalytic mechanism. Except that a small quantity of scavenger is introduced to the reaction system, the experimental procedure is quite identical to the photodegradation experiment.
To assess the photocatalytic stability of the AT2 nanocomposite, cycle photocatalytic studies were carried out using the aforementioned procedures. The catalyst was gathered after every cycle of the photocatalytic procedure, cleaned with deionized water, and then re-dispersed into a new RhB solution for the following cycle. Trapping agent studies were used to investigate the photocatalytic mechanism. Except that a small quantity of scavenger is introduced to the reaction system, the experimental procedure is quite identical to the photodegradation experiment.
9
Anthocyanin Measurement Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The anthocyanin measurement was conducted according to a previous report [14 (link)]. Approximately 0.5 g fresh tissue was ground in liquid nitrogen, and added to 5 mL extraction solution (methanol: H2O: formic acid: trifluoroacetic acid, 70:27:2:1), and kept at 4 °C in the dark. After 12 h, the supernatant was collected by centrifugation at 13,000 rpm for 10 min at 4 °C. Then, the absorbance was measured at 530 nm and 657 nm by UV spectrophotometer (UV-1800PC, MAPADA, Shanghai, China). The formula for the calculation of the anthocyanins content is as follows: Total anthocyanins = [A530 − (0.25 × A657)]/M, where A530 and A657 are the absorbance at the indicated wavelengths, and M is the fresh tissue weight (g).
10
Cumulative Release of Antimicrobial Peptides
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
AMPs release ratio from hydrogels was calculated using a previously published method with slight modifications [24 (link)]. Briefly, drug loaded hydrogels were introduced into a 30 mL deionized water. Sample aliquots of 3 mL were withdrawn at regular time intervals of 2, 4, 6, 8, 12, 14 and 24 h. An equal volume of deionized water was added at each step of sample withdrawn to keep the release medium volume constant throughout the experiment. AMPs concentration released as a function of time was measured using an ultravioletray spectrophotometer (MAPADA UV-1800PC, China) at λmax 257 nm. The concentration of AMPs is calculated by comparing optical density of the samples with a standard curve (Supplementary Fig. 1 ). AMPs concentration released thus determined was plotted as cumulative drug release vs. time.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!