Bca protein assay

Manufactured by Santa Cruz Biotechnology

Sourced in United States

The BCA protein assay is a colorimetric detection and quantification method used to determine the total protein concentration in a sample. It is based on the reduction of copper ions by proteins in an alkaline medium, which produces a purple-colored complex that can be measured spectrophotometrically.

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Lab products found in correlation

Gapdh antibody, by Merck Group (1 mentions) β actin antibody, by Merck Group (1 mentions) Enhanced chemiluminescence detection system, by Bio-Rad (1 mentions) Pvdf membrane, by Roche (1 mentions) Anti β tubulin, by Santa Cruz Biotechnology (1 mentions) Ecl detection system, by GE Healthcare (1 mentions) Odyssey infrared imager, by LI COR (1 mentions) Goat anti mouse irdye 800, by LI COR (1 mentions) Goat anti rabbit irdye 680, by LI COR (1 mentions) Anti vsv g, by Abcam (1 mentions) Anti gapdh, by Beyotime (1 mentions) Protein g plus agarose bead, by Santa Cruz Biotechnology (1 mentions) Anti flag, by Merck Group (1 mentions) Anti myc, by Santa Cruz Biotechnology (1 mentions) Quantity one software, by Bio-Rad (1 mentions) Iseq00010, by Merck Group (1 mentions) Peroxidase linked goat anti rabbit igg, by Bioworld Technology (1 mentions) Ecl detection system, by Vazyme (1 mentions)

Close spelling variants of this product

BCA protein assay kit (29 citations) BCA assay (7 citations)

5 protocols using bca protein assay

Examining Involucrin Expression in HaCaT Cells

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In order to examine the expression of involucrin in the HaCaT cell line, cells were washed with PBS, then collected and solubilized in RIPA buffer on ice for 30 min. The lysate was centrifuged at 12,000 rpm for 15 min at 4 °C to remove cellular debris and the supernatant protein concentration was determined by the BCA protein assay (Santa Cruz Biotechnology, Santa Cruz, CA, USA). The proteins were then added to 5 × loading buffer and incubated at 95 °C for 5 min before being subjected to standard western blotting.
The primary antibody against involucrin (Abcam, Cambridge, UK) were diluted at 1:500, ATP5B antibody (Abcam, Cambridge, UK) was diluted at 1:400, β-actin antibody (Sigma, Germany) was diluted at 1:10,000, and GAPDH antibody (Sigma, Germany) was diluted at 1:5000. Immunolabeled proteins were then visualized using a goat anti-mouse antibody linked to horseradish peroxidase (1:10,000, Sigma, Germany) and an enhanced chemiluminescence detection system (Bio-Rad, Hercules, CA, USA). Protein levels in each lane were normalized to that of β-actin or GAPDH.

Xiaoyun X., Chaofei H., Weiqi Z., Chen C., Lixia L., Queping L., Cong P., Shuang Z., Juan S, & Xiang C. (2017). Possible Involvement of F1F0-ATP synthase and Intracellular ATP in Keratinocyte Differentiation in normal skin and skin lesions. Scientific Reports, 7, 42672.

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Testis Protein Extraction and Western Blot

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Proteins from the samples were extracted as described previously [20 (link)]. Testis samples from both groups were homogenized in ice-cold RIPA buffer (25 mM Tris/HCl (pH 7.6), 1% sodium deoxycholate, 150 mM NaCl, 0.1% SDS, 1% Nonidet-P40, and 0.05 mM PMSF) and centrifuged at 15,000 rpm for 10 min at 4 °C. Subsequently, the concentration of total protein was evaluated using a BCA protein assay (Santa Cruz Biotechnology, sc-202389). Samples (40 μg protein per lane) were subjected to 10% SDS-PAGE gel electrophoresis and then transferred to PVDF membranes (Roche). Membranes were blocked with 5% milk powder in 1 × 9 phosphate-buffered saline (PBS) and 0.1% Tween 20 for 60 min, washed with PBS/Tween, and incubated overnight at 4 °C with primary antibodies, including anti-ESRα and BOLL (1:500 dilution; Abcam, Cambridge, UK) and anti-β-tubulin (1:1000 dilution; Santa Cruz Biotechnology). Membranes were then incubated for 2 h with an anti-rabbit secondary antibody, and bands were imaged using an ECL detection system (GE Healthcare Bio-Sciences, Pittsburgh, PA, USA). Finally, photographs were taken after exposure to the X-ray films.

Kalwar Q., Chu M., Ahmad A.A., Xiong L., Zhang Y., Ding X, & Yan P. (2021). Expressional Profiling of TEX11, ESRα and BOLL Genes in Yak under Different Feeding Conditions. Biology, 10(8), 731.

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Western Blot Protein Detection Protocol

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Cells were lysed with RIPA lysis buffer (Cat. No. sc-24948A; Santa Cruz, USA) and total protein concentrations were determined by the BCA protein assay (Cat. No. 23225; Pierce Rockford, IL, USA). Equal concentrations of harvested protein were electrophoresed on 10% (w/v) sodium dodecyl sulfate (SDS) polyacrylamide gel and transferred onto a 0.45 μm PVDF membrane (Cat. No. 03010040001; Millipore, Billerica MA, USA) which was activated in methanol. The primary antibodies used to detect the target proteins were as following: anti-VSV-G was purchased from Abcam (Cat. No. ab183497; Cambridge, UK). Anti-RSV-G and anti-GFP were both purchased from Sino Biological (Cat. No. 13029-T52; Cat. No. 13105-R208; Beijing, China). Anti-FOLR1 and anti-APOBEC3B were purchased from ABclonal (Cat. No. A15672; Cat. No. A9010; Wuhan, China). Anti-GAPDH was purchased from Beyotime (Cat. No. AF0006; Shanghai, China). IRDye 680 goat-anti-rabbit and IRDye 800 goat-anti-mouse were purchased from Li-COR (Cat. No. 926-68071; Cat. No. 926-32216; Lincoln, NE, USA), and co-incubated with the membrane. Then they were visualized by a Li-COR Odyssey Infrared Imager. The protein bands were quantified using ImageJ (National Institutes of Health and the Laboratory for Optical and Computational Instrumentation, university of Wisconsin, America).

Wu J., Han Y., Lyu R., Zhang F., Jiang N., Tao H., You Q., Zhang R., Yuan M., Nawaz W., Chen D, & Wu Z. (2023). FOLR1-induced folate deficiency reduces viral replication via modulating APOBEC3 family expression. Virologica Sinica, 38(3), 409-418.

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Immunoprecipitation of Myc-tagged HMGB1 and Sp1-Flag

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293FT cells were transfected with the Myc-pCMV-hHmgb1 and pCMV-Sp1-Flag plasmids (purchased from GeneChem, Shanghai, China). The cells were harvested 36 h later and incubated in IP lysis buffer on ice for 30 min. The protein concentration of each samples was determined using the BCA protein assay (Santa Cruz Biotech, Santa Cruz, CA, USA). Cell lysates were centrifuged at 6000 rpm for 4 min to pellet cellular debris. The supernatant were then incubated with anti-Flag (Sigma-Aldrich Corp., St. Louis, MO), anti-Myc (Santa Cruz Biotech, Santa Cruz, CA, USA) or control IgG at 4°C for 2 hours. Protein G PLUS-agarose beads were then added (Santa Cruz Biotech, Santa Cruz, CA), and the mixtures were rotated overnight at 4°C. After washing thoroughly, the beads were incubated at 95°C for 5min in approximately 40 μl lysis buffer plus 10 μl 5 × loading buffer. The protein samples were subjected to western blot analysis.

Li Q., Li J., Wen T., Zeng W., Peng C., Yan S., Tan J., Yang K., Liu S., Guo A., Zhang C., Su J., Jiang M., Liu Z., Zhou H, & Chen X. (2014). Overexpression of HMGB1 in melanoma predicts patient survival and suppression of HMGB1 induces cell cycle arrest and senescence in association with p21 (Waf1/Cip1) up-regulation via a p53-independent, Sp1-dependent pathway. Oncotarget, 5(15), 6387-6403.

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Testis Protein Extraction and Western Blot

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Samples of the testis in each group were homogenized in ice-cold RIPA buffer (25 mM Tris/HCl (pH 7.6), 150 mM NaCl, 1% sodium deoxycholate, 1% Nonidet-P40, 0.1% SDS, 0.05 mM PMSF), and centrifuged at 15,000 g for 10 min at 4°C. Then, the total protein concentration was determined with a BCA protein assay (Santa Cruz, sc-202389). Samples (40 μg protein per lane) were subjected to electrophoresis on a 10% SDS-PAGE gel and then transferred onto PVDF membranes (Millipore, ISEQ00010). After nonspecific blocking in 5% nonfat milk, the membranes were incubated with an anti-LC3B (1:1000 dilution) antibody (Abcam, ab48394) overnight at 4°C. After washing with TBST, the membranes were incubated with peroxidase-linked goat anti-rabbit IgG (1:5000, Bioworld Technology Inc., BS13278) for 2 h. Following incubation, the bound antibodies were visualized by using the ECL detection system (Vazyme Biotech, E411-04). Immunoreactive bands were quantified with Quantity One software (Bio-Rad Laboratories).

Huang Y., Yang P., Liu T., Chen H., Chu X., Ahmad N., Zhang Q., Li Q., Hu L., Liu Y, & Chen Q. (2016). Subcellular Evidence for Biogenesis of Autophagosomal Membrane during Spermiogenesis In vivo. Frontiers in Physiology, 7, 470.

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