Log In Sign up
The largest database of trusted experimental protocols

Bca protein reagent

Manufactured by Thermo Fisher Scientific
Visit Supplier
Sourced in United States

The BCA protein reagent is a laboratory product used to quantify protein concentrations in samples. It is a colorimetric detection method that measures the absorbance of a purple-colored reaction product formed by the chelation of two molecules of bicinchoninic acid with one cuprous ion, which is proportional to the amount of protein present in the sample.

Automatically generated - may contain errors

Lab products found in correlation

Sds polyacrylamide gel, by Bio-Rad (5 mentions) Anti β actin, by Merck Group (4 mentions) Enhanced chemiluminescence western blotting detection reagent, by Cytiva (3 mentions) Cell signaling buffer, by Cell Signaling Technology (3 mentions) Ecl reagent, by PerkinElmer (3 mentions) Ripa buffer, by Thermo Fisher Scientific (2 mentions) Anti bcl xl, by Cell Signaling Technology (2 mentions) Anti active caspase 3, by Abcam (2 mentions) Tissue lysis extraction reagent, by Merck Group (1 mentions) Protease inhibitor cocktail, by Roche (1 mentions) Anti glyceraldehyde 3 phosphate dehydrogenase gapdh, by Santa Cruz Biotechnology (1 mentions) Recombinant human cxcl12, by Thermo Fisher Scientific (1 mentions) Anti actin, by Santa Cruz Biotechnology (1 mentions) Tcrαβ fitc, by BD (1 mentions) Cd20 apc, by BD (1 mentions) Il 21, by Thermo Fisher Scientific (1 mentions) Anti erk1 2, by Cell Signaling Technology (1 mentions) Cd38 percp cy5, by BD (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

Pierce BCA Protein Assay Reagents A and B Pierce BCA Protein Assay Reagent A BCA Protein Assay Reagent Kit BCA protein reagent kit BCA Protein Quanti cation Reagent Kit Bicinchoninic acid (BCA) protein assay reagent kit Micro-BCA protein reagent Pierce BCA Protein Assay Reagent Kit Micro BCA Protein Assay Reagent Kit Bicinchoninic acid (BCA) protein assay reagent

17 protocols using bca protein reagent

1

Protein Extraction from Prostatic Tissue

Check if the same lab product or an alternative is used in the 5 most similar protocols
Prostatic tissue was homogenized (1/10 w/v) in tissue lysis/extraction reagent (Sigma-Aldrich, St. Louis, MO, USA) containing protease inhibitor cocktail (Roche, Mannheim, Germany) using a homogenizer. Homogenates were centrifuged at 15,500× g for 20 min at 4 °C. Total protein concentrations in the supernatant fractions were measured using BCA protein reagent (Thermo Fisher Scientific, Waltham, MA, USA).
Xin Q., Kwon M.J., Lee J.W., Kim K.S., Chen H., Campos M.G., Tundis R., Cui C.B., Cho Y.H, & Cao H. (2019). Gamma Irradiated Rhodiola sachalinensis Extract Ameliorates Testosterone-Induced Benign Prostatic Hyperplasia by Downregulating 5-Alpha Reductase and Restoring Testosterone in Rats. Molecules, 24(21), 3981.
+ Open protocol
+ Expand
2

Immune Cell Activation by CXCL12 and IL-21

Check if the same lab product or an alternative is used in the 5 most similar protocols
Recombinant human CXCL12 and IL-21 were purchased from Peprotech (Rocky Hill, NJ). Peptidoglycan (PGN) poly (I:C), LPS, and R848 were purchased from Sigma-Aldrich (Poole, Dorset, UK). Human CpG-B DNA was purchased from Hycult Biotech (Uden, Netherlands). Flagellin was provided by Dr. Myoung Ho Jang (Osaka University). RPMI 1640 medium and fetal bovine serum (FBS) were purchased from Gibco BRL (Eggenstein, Germany). Anti-ERK1/2 and anti-human phospho-AKT1/2/3 monoclonal antibodies (Ab) were obtained from Cell Signaling Technology. Anti-ß-actin and anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) Abs were obtained from Santa Cruz (Paso Robles, CA). Anti-RGS1 Ab was purchased from Novus Biologicals (Littleton, CO). The anti-CXC chemokine receptors (CXCR) 4-APC, TCRαβ-FITC, CD38-PerCP-Cy5.5, and CD20-APC were purchased from BD Biosciences (Heidelberg, Germany). The BCA protein reagent was purchased from Thermo Scientific (Hudson, NH). Interleukin (IL)-2 and IL-4 were provided by Prof. Jongseon Choe (College of Medicine, Kangwon National University).
Pak H.K., Gil M., Lee Y., Lee H., Lee A.N., Roh J, & Park C.S. (2015). Regulator of G Protein Signaling 1 Suppresses CXCL12-Mediated Migration and AKT Activation in RPMI 8226 Human Plasmacytoma Cells and Plasmablasts. PLoS ONE, 10(4), e0124793.
+ Open protocol
+ Expand
3

Cell Lysis and Western Blot Analysis

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were lysed using boiling lysis buffer (1% SDS, 10 mM Tris.HCl, pH7.4). Protein concentrations were measured using BCA protein reagent (Thermo Scientific) and equal amounts of protein were separated on 10% or gradient polyacrylamide gels and transferred onto PVDF membranes [69 (link), 81 (link)]. The following antibodies were used: rabbit antibodies against phospho-pS6K(T389), phospho-S6(S235/236) and phospho-S6(S240/244), S6K, phospho-4EBP1(T37/46), phospho-AKT(S473) and phospho-AKT(T308), AKT and mouse anti-S6 – from Cell Signaling Technology (Danvers, MA); mouse monoclonal antibodies against cyclin D1 and rabbit anti-actin were from Santa Cruz Biotechnology (Paso Robles, CA) and Sigma-Aldrich (St. Louis, MO), respectively.
Leontieva O.V, & Blagosklonny M.V. (2016). Gerosuppression by pan-mTOR inhibitors. Aging (Albany NY), 8(12), 3535-3549.
+ Open protocol
+ Expand
4

Femur Decalcification and Protein Extraction

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
The mouse femurs were dissected and decalcified by EDTA before protein extraction [14 (link)]. The decalcified bone samples were chopped into small pieces and homogenized with TissueLyser 5mm stainless steel beads (Qiagen, Redwood City, CA) in the RIPA lysis buffer containing the proteinase inhibitor cocktail (ThermoFisher Scientific, Waltham, MA). Protein concentrations were measured with the BCA protein reagent (ThermoFisher Scientific). The p65 (D14E12) and phospho-p65 (93H1) were stained by the antibodies from Cell Signaling Technology (Danvers, MA). The horseradish peroxidase-conjugated secondary antibody (Cell Signaling Technology) was used and the signals were detected by adding the enhanced chemiluminescence western blotting detection reagents (ThermoFisher Scientific). The quantification of western blotting results was done by Image Lab statistic software (Bio-Rad). The images were quantified by using ImageJ.
Lin T., Pajarinen J., Kohno Y., Nabeshima A., Lu L., Nathan K., Yao Z., Wu J.Y, & Goodman S. (2019). Increased NF-kB activity in osteoprogenitor-lineage cells impairs the balance of bone versus fat in the marrow of skeletally mature mice. Regenerative engineering and translational medicine, 6, 69-77.
+ Open protocol
+ Expand
5

Quantification of Cyanobacterial Biomass

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Cell dry weight concentrations were measured directly as ash- free dry weight (AFDW) (Pinchuk et al., 2010 (link)) and compared in a standard curve to the indirect OD730 measurements obtained using a Genesys 20 visible spectrophotometer (Thermo Scientific, Rockford, IL). Protein was quantitated using BCA protein reagent (Thermo Scientific). Chl a and phycocyanin concentrations were estimated using a previously described method that corrects the effect of scatter on absorbance from whole-cell suspensions (see Supplemental Methodologies) (Myers, 1980 ; Burns et al., 2006 (link)). Dissolved O2 concentration in the reactor was measured with a Clark O2 electrode (InPro® 6800Series, Mettler Toledo International Inc., Columbus, OH). O2 production rates as a function of “white light” irradiance (tungsten incandescent) were measured inside an oxygraph chamber (Hansatech, Norfolk, UK).
Bernstein H.C., Konopka A., Melnicki M.R., Hill E.A., Kucek L.A., Zhang S., Shen G., Bryant D.A, & Beliaev A.S. (2014). Effect of mono- and dichromatic light quality on growth rates and photosynthetic performance of Synechococcus sp. PCC 7002. Frontiers in Microbiology, 5, 488.
+ Open protocol
+ Expand
6

Quantitative Analysis of Superoxide

Check if the same lab product or an alternative is used in the 5 most similar protocols
For detection of superoxide, cells were washed twice with DPBS and incubated with Hank’s Balanced Salt Solution (HBSS) containing 10 μM dihydroethidium (DHE) for 60 mins. Media were collected and centrifuged at 3000 rpm for 5 mins. Fluorescence analysis of the oxidation product 2-hydroxyethidium (EOH) was performed on a Beckman-Coulter HPLC system. Samples were separated on a Synergi-Fusion column (250×4.6 mm, Phenomenex) using a gradient elution of 10% v/v acetonitrile, 0.1% trifluoracetic acid solution (TFA) as the mobile phase A and 100% acetonitrile as mobile phase B with a flow rate set at 1.00 ml/min. Runs began at 100% phase A. Phase B was increased linearly from 0% to 42% in 25 mins, then reduced to 0% during 5 mins and kept at 0% for 5 minutes. The UV detector was set at 350 nm to monitor the elution of DHE. Fluorescence detection was measured using 480 nm (excitation) and 580 nm (emission). Authentic DHE and EOH standard solutions were injected onto the HPLC to determine the retention time of the corresponding peaks. All procedures were performed in the dark or under indirect light. Quantification was performed by normalizing the integrated peak areas with protein concentrations measured using the BCA Protein Reagent (Thermo Scientific).
Bera S., Weinberg F., Ekoue D.N., Ansenberger-Fricano K., Mao M., Bonini M.G, & Diamond A.M. (2014). Natural allelic variations in glutathione peroxidase-1 affect its subcellular localization and function*. Cancer research, 74(18), 5118-5126.
+ Open protocol
+ Expand
7

Fluorometric Assay for MMP Activity

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Leukemia cell lines were lysed with RIPA buffer (Sigma) following the manufacturer’s instructions. Protein concentrations of the lysates were measured with the BCA protein reagent (Pierce Chemical). 20 µl of lysate was added to 80 µl MMP buffer (10 mM Tris pH 7.5, 150 mM NaCl, 10 mMCaCl2, 0.05% Triton X-100). Fluorogenic peptide substrate IX (R&D Systems) was added to a final concentration of 10 µM. Samples were read in a fluorescent plate reader (excitation 320 nm, emission 405 nm) every 10 min for 70 minutes. Relative fluorescence units were normalized to protein concentration of the lysates [20] (link).
Hsu Y.C., Mildenstein K., Hunter K., Tkachenko O, & Mullen C.A. (2014). Acute Lymphoid Leukemia Cells with Greater Stem Cell Antigen-1 (Ly6a/Sca-1) Expression Exhibit Higher Levels of Metalloproteinase Activity and Are More Aggressive In Vivo. PLoS ONE, 9(2), e88966.
+ Open protocol
+ Expand
8

Western Blot Analysis of Androgen Receptor

Check if the same lab product or an alternative is used in the 5 most similar protocols
2.5×106 cells (per 100-mm dish) were washed with 1x PBS and scraped into a lysis buffer. Protein concentrations were measured with the BCA protein reagent (Pierce Chemical, Rockford, IL). Approximately 50 μg of protein/lane were loaded and run on a 10% polyacrylamide gel with a Tris/glycine running buffer system and then transferred onto a polyvinylidene difluoride membrane. The blots were probed with primary anti-AR (N-20), anti-PSA (C-19) antibodies with dilutions of 1:500 to 1:1,000 and incubated at room temperature for 2 hrs. The secondary antibody [rabbit anti-goat IgG, 1:5,000 dilution (Santa Cruz Biotechnology) or rabbit anti-mouse IgG, 1:5,000 dilution (Pierce Chemical, Rockford, IL)] was used at room temperature for 1 hr. Immunoblot analysis was performed with horseradish peroxidase- conjugated anti-rabbit or anti-mouse IgG antibodies using enhanced chemiluminescence Western blotting detection reagents (Amersham Biosciences). The quantification of Western blotting results was done by Image Lab statistic software (Bio-red).
Xu D., Chen Q., Liu Y, & Wen X. (2017). Baicalein suppresses the androgen receptor (AR)-mediated prostate cancer progression via inhibiting the AR N-C dimerization and AR-coactivators interaction. Oncotarget, 8(62), 105561-105573.
+ Open protocol
+ Expand
9

Protein Quantification and Western Blot

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cell lysates were harvested and 20-100 ug of protein were resolved by 12% SDS-PAGE gel after measuring protein concentration using the BCA protein reagent (Pierce Chemical, Rockford, IL, USA) and then transferred to nitrocellulose membranes. The membranes were blocked with 5% nonfat milk for 1 h at room temperature and then incubated with primary antibodies overnight at 4°C, followed by incubation with HRP-conjugated anti-rabbit/ mouse/ goat IgG for 1 h at room temperature. Detection was performed using enhanced chemiluminescence (ECL) detection reagent (Thermo).
Wang H., Huang F., Zhang Z., Wang P., Luo Y., Li H., Li N., Wang J., Zhou J., Wang Y, & Li S. (2019). Feedback Activation of SGK3 and AKT Contributes to Rapamycin Resistance by Reactivating mTORC1/4EBP1 Axis via TSC2 in Breast Cancer. International Journal of Biological Sciences, 15(5), 929-941.
+ Open protocol
+ Expand
10

Western Blot Protein Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols
The cells were washed with 1xPBS and scraped into a lysis buffer containing the proteinase inhibitor cocktail (Roche). Protein concentrations were measured with the BCA protein reagent (Pierce Chemical, Rockford, IL). Approximately 50 μg of protein/lane were loaded and run on the polyacrylamide gel with a Tris/glycine running buffer system and then transferred onto a polyvinylidene difluoride membrane. The primary antibodies with dilutions of 1:500 to 1:1,000 were added and then incubated overnight in the cold room with vigorous shaking. The horseradish peroxidase-conjugated secondary antibody (Pierce Chemical, Rockford, IL) was added and the signals were detected by adding the enhanced chemiluminescence Western blotting detection reagents (Amersham Biosciences, Piscataway, NJ).
Ding X., Yang D.R., Xia L., Chen B., Yu S., Niu Y., Wang M., Li G, & Chang C. (2015). Targeting TR4 nuclear receptor suppresses prostate cancer invasion via reduction of infiltrating macrophages with alteration of the TIMP-1/MMP2/MMP9 signals. Molecular Cancer, 14(1), 16.
+ Open protocol
+ Expand

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

Sign up now

Revolutionizing how scientists
search and build protocols!