Bca protein assay kit

Manufactured by Wuhan Servicebio Technology

Sourced in China

The BCA protein assay kit is a colorimetric detection and quantitation assay for determining the total protein content of a sample. It utilizes bicinchoninic acid (BCA) as the detection reagent, which changes color in proportion to the amount of protein present in the sample. The kit provides a simple and accurate method for measuring protein concentration, making it a useful tool for various biochemical and analytical applications.

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Lab products found in correlation

Pvdf membrane, by Merck Group (8 mentions) Ripa lysis buffer, by Wuhan Servicebio Technology (6 mentions) Ripa buffer, by Wuhan Servicebio Technology (6 mentions) β actin, by Wuhan Servicebio Technology (3 mentions) Ripa lysate, by Beyotime (2 mentions) Ipvh00010, by Merck Group (2 mentions) Anti atcb, by Immunoway (2 mentions) Electrophoresis chamber, by Merck Group (2 mentions) Halt protease and phosphatase inhibitor cocktail, by Wuhan Servicebio Technology (2 mentions) Anti psmb3, by Solarbio (2 mentions) Anti psmb6, by Solarbio (2 mentions) Anti psma4, by Solarbio (2 mentions) Anti rabbit igg hrp conjugated secondary antibody, by ABclonal (2 mentions) Gapdh, by Abcam (2 mentions) Pvdf membrane, by Wuhan Servicebio Technology (2 mentions) Hrp labeled goat anti rabbit igg, by Wuhan Servicebio Technology (1 mentions) Alpha imager, by Bio-Techne (1 mentions) Sds page gel, by Beyotime (1 mentions) Beyoecl moon chemiluminescence system, by Beyotime (1 mentions) Ab201688, by Abcam (1 mentions)

Spelling variants of this product

BCA kit (39 citations) BCA protein assay (6 citations) BCA assay kit (4 citations)

53 protocols using bca protein assay kit

Isolation and Characterization of Mesenchymal Stem Cell-Derived Exosomes

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The typical ultracentrifugation method was used to extract and collect the exosomes from the supernatant of the GMSCs culture. When the GMSCs (P3–P5) had achieved a confluence of 80–85%, the medium was replaced by fresh DMEM/F12 containing 10% exosome-free FBS, and then the cells were further cultures for 48 h. The supernatant was collected and centrifuged at 500g for 10 min, 2000g for 20 min, 10 000g for 30 min sequentially to eliminate any dead cells and cell debris. Clear supernatants were passed through 0.22 μm filters and transferred into 15 mL Amicon Ultra-30kd ultracentrifugation tubes, for ultracentrifugation at 100 000g for 60 min twice. The final precipitates from the supernatant were resuspended with PBS and stored at −80 °C.
Transmission electron microscopy (TEM, HITACHI) was utilized to observe the morphology of the GMSCs-Exo. Nanoparticle tracking analysis (NTA, IZON) was used to measure the size of the exosomes and analyze the size distribution. The concentration of GMSCs-Exo was determined by the BCA protein assay kit (Servicebio), while the exosomal markers CD63 and Tsg101 were detected by western blot analysis.

Liu Z., Yang S., Li X., Wang S., Zhang T., Huo N., Duan R., Shi Q., Zhang J, & Xu J. (2022). Local transplantation of GMSC-derived exosomes to promote vascularized diabetic wound healing by regulating the Wnt/β-catenin pathways. Nanoscale Advances, 5(3), 916-926.

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Western Blot Protein Quantification

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The livers were lysed and centrifuged to gain total protein extracts. The bicinchoninic acid (BCA) protein assay kit (Servicebio, China) was used to measure protein concentrations. After separation by electro phoresis on 10% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) gels and transmembrane onto polyvinylidene difluoride (PVDF) membrane, the samples were blocked with 10% skimmed milk. Then, membranes were incubated with primary antibodies at 4°C for 12 h. Subsequently, membranes were washed and then incubated with secondary antibodies at room temperature for 30 min followed by visualization using the Electrochemiluminescence system. Representative blots were chosen from three independent experiments. Optical density value of western blots was performed with the use of Alpha software. Protein levels were normalized against those of corresponding β-actin.

Lu Y.H., Hong Y., Zhang T.Y., Chen Y.X., Wei Z.J, & Gao C.Y. (2022). Rosmarinic acid exerts anti-inflammatory effect and relieves oxidative stress via Nrf2 activation in carbon tetrachloride-induced liver damage. Food & Nutrition Research, 66, 10.29219/fnr.v66.8359.

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Quantification of NF-κB p65 Protein in Bursa

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The western blot method was conducted to detect the protein expression levels of bursa of Fabricius. The total protein, nuclear protein and cytoplasmic protein was extracted from bursa of Fabricius tissues and the protein concentration was determined using the BCA protein assay kit (Servicebio Technology, Wuhan, China). And then the protein was assessed by SDS-polyacrylamide gel electrophoresis under reducing conditions on 10% gels. Subsequently, it was transferred to nitrocellulose membranes using a tank transfer at 200 mA in Tris-glycine buffer containing 20% methanol for 90 min at 4°C, and then put the membranes in 5% skim milk for block at 37°C 1 h. The membranes were consistently incubated overnight at 4°C with diluted primary antibody that a rabbit polyclonal antibody against nuclear factor-kappa B (NF-κB) p65 (GB11997, 65 kDa, Servicebio, Wuhan, China). The diluted concentration of primary antibody was 1:1000. The HRP-labeled goat anti-rabbit IgG (1:3000, Servicebio Technology, Wuhan, China) was used as the secondary antibody. The β-actin content was analyzed as the loading control with rabbit IgG (1:3000) polyclonal antibodies. The proteins bands detected and analyzed using Alpha Imager (Alpha Innotech, CA). The relative protein expression levels were calculated as target protein/β-actin.

Liu W.C., Ou B.H., Liang Z.L., Zhang R, & Zhao Z.H. (2021). Algae-derived polysaccharides supplementation ameliorates heat stress-induced impairment of bursa of Fabricius via modulating NF-κB signaling pathway in broilers. Poultry Science, 100(8), 101139.

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Comprehensive Western Blot Analysis

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Western blot was conducted as previously described [23 (link)]. Briefly, protein concentration in the supernatant of cells was determined using a BCA Protein Assay Kit (Servicebio). Proteins (30 μg) were separated on an SDS-PAGE gel (Beyotime) and transferred to PVDF membranes. Subsequently, the membranes were incubated overnight with primary antibodies against anti-Bcl-2, anti-Bax, anti-caspase 3, anti-caspase 9, anti-Cytochrome c (Cyt-c), anti-apoptosis-inducing factor (AIF), anti-Smac/DIABLO, anti-Tnmd, anti-MMP-1, anti-Col I, anti-Col III, anti-Dynamin-related protein 1 (Drp1), anti-Mitofusin 2 (Mfn2), and anti-β-actin. The membrane was then incubated with horseradish peroxidase (HRP)-conjugated secondary antibody for 1 h at 22–26 °C. Protein bands were visualized using the BeyoECL Moon chemiluminescence system (Beyotime). All antibody information and dilutions are listed in Additional file 5: Table S2.

Wei B., Ji M., Lin Y., Wang S., Liu Y., Geng R., Hu X., Xu L., Li Z., Zhang W, & Lu J. (2023). Mitochondrial transfer from bone mesenchymal stem cells protects against tendinopathy both in vitro and in vivo. Stem Cell Research & Therapy, 14, 104.

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Protein Extraction and Western Blot Analysis

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The total cellular protein was extracted using radioimmunoprecipitation assay (RIPA) lysis buffer. The extracted protein concentration was quantified using a BCA Protein Assay Kit (Servicebio, Wuhan, China). The protein was separated using SDS-PAGE. Target proteins were transferred onto the polyvinylidene fluoride membranes. 5% skim milk powder was used to block the transferred membranes, which were then incubated with anti-CSDE1 (Abcam, ab201688) and anti-GAPDH (Abcam, ab181603) primary antibodies at 4°C overnight. After rinsing with Tris-Buffered Saline with Tween® 20, the membranes were incubated with secondary antibodies (BOSTER, BA1056) at 25°C for 1 hour. Finally, target protein signals were visualized after enhanced chemiluminescence.

Yu J., Zhang X., He R., Yang X., Hu J., Tan F., Yao J., Lei X, & Wen G. (2022). LINC01234 Accelerates the Progression of Breast Cancer via the miR-525-5p/Cold Shock Domain-Containing E1 Axis. Disease Markers, 2022, 6899777.

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Radiation-Induced DNA Damage Signaling

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Cells pretreated with nimotuzumab received irradiation as described above. Cytoplasmic and nuclear extracts were prepared according to the directions for using the nuclear and cytoplasmic extraction kit (Beyotime, Nanjing, Jiangsu, People’s Republic of China) 6 hours after irradiation. Protein concentration was determined by the BCA protein assay kit (Servicebio, Wuhan, Hubei, People’s Republic of China). Protein lysates were then separated on an 8% or 12% SDS-PAGE gel depending on the protein molecular weight and transferred to PVDF membranes (Millipore, Billerica, MA, USA). Immunoblots were blocked in 5% protease-free bovine serum albumin for 1 hour and probed with γ-H2AX (Abcam, Cambridge, UK), EGFR and p-EGFR (Cell Signaling Technology, Beverly, MA, USA), DNA-PK (Cell Signaling Technology, Beverly, MA, USA), p-DNA-PK (Epitomics, Burlingame, CA, USA), as well as GAPDH and Lamin B1 (Goodhere, Hangzhou, Zhejiang, People’s Republic of China) antibodies. The membranes were continually incubated with appropriate horseradish peroxidase secondary antibodies (Invitrogen, Carlsbad, NM, USA) after washing, and then we detected protein with a chemiluminescence kit (Invitrogen) and visualized the bands on an X-ray film. GAPDH and Lamin B1 were used as an internal control for cytoplasmic and nuclear proteins, respectively, to balance equal loading.

Teng K., Zhang Y., Hu X., Ding Y., Gong R, & Liu L. (2015). Nimotuzumab enhances radiation sensitivity of NSCLC H292 cells in vitro by blocking epidermal growth factor receptor nuclear translocation and inhibiting radiation-induced DNA damage repair. OncoTargets and therapy, 8, 809-818.

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Western Blot Analysis of Protein Expression

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RIPA lysate (Beyotime, Shanghai, China) was used to extract total proteins from the diversely treated HMCs and renal tissues of mice for western blot analysis. A bicinchoninic acid (BCA) protein assay kit (G2026, Servicebio, China) was used to determine protein concentration. Equivalent quantities of the protein samples were separated by 4%-12% SurePAGE (M00652, GenScript, USA), transferred to a PVDF membrane, and blocked with 5% BSA or dry milk in TBST for 1 h at 23°C ± 2°C. The primary antibodies (1:1000 dilution) were incubated 12-16 h at 4°C, the membranes were washed 3 times with TBST, and incubated for 1 h at 23°C ± 2°C with horseradish peroxidase-conjugated secondary antibodies (1:1000 dilution). The membranes were then incubated with enhanced chemiluminescence (ECL) reagent and imaged using the Tanon 5,200 luminescent imaging workstation. The ImageJ software was used to analyze the images. A strip solution washed membranes which needed to be imaged again. The representative experimental results were repeated three times.

Wang Y., Liu Y., Chen S., Li F., Wu Y., Xie X., Zhang N., Zeng C., Bai L., Dai M., Zhang L, & Wang X. (2023). The protective mechanism of Dehydromiltirone in diabetic kidney disease is revealed through network pharmacology and experimental validation. Frontiers in Pharmacology, 14, 1201296.

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Cerebral Cortex and Gastric ChAT Expression

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A Western blot analysis was performed to assess the expression of ChAT protein in the cerebral cortex on the infarcted side and in gastric tissues. The total protein concentration in each group was determined using the BCA Protein Assay Kit (G2026-200T, Servicebio). Subsequently, 10 μg of each sample was loaded onto a 10 % SDS-PAGE gel for electrophoresis. Following electrophoresis, the proteins were transferred onto PVDF membranes (IPVH00010, Millipore) and blocked with 5 % skim milk for 1 h at room temperature. The membranes were then incubated overnight at 4 °C on a shaker with primary antibodies, including ChAT (1:1000, ab181023, Abcam), α7nAchR (1:1000, ab216485, Abcam), and β-actin (1:1000, GB15003, Servicebio). On the following day, the membranes were incubated with a fluorescent secondary antibody (1:10000, SA5-35571, Invitrogen) and visualized using an Odyssey imager.

Jin Z., Shen Z., Yan S., Chen G., Yin Y., Zhang Y, & Wu X. (2024). Electroacupuncture ameliorates gastrointestinal dysfunction by modulating DMV cholinergic efferent signals to drive the vagus nerve in p-MCAO rats. Heliyon, 10(8), e29426.

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Protein Expression Analysis Protocol

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Tissue samples were used to extract total protein using the RIPA lysis buffer (G2002, Servicebio, Wuhan, China). The protein concentrations were determined using the BCA protein assay kit (G2026, Servicebio, Wuhan). Electrophoresis was conducted on a 10% sodium dodecyl sulfate-polyacrylamide gel, and the resulting proteins were subsequently transferred onto polyvinylidene fluoride (PVDF) membranes. The membranes were then incubated with primary antibodies against AQP1 (1:1000), SOD1 (1:1000), and MUC2 (1:1000) overnight at 4 °C. HRP-labeled secondary antibodies were incubated at room temperature for 2 h. The PVDF membranes were washed 5 times with Tris-buffered saline tween (TBST) and developed using an electrochemiluminescence reagent.

Sun C., Wang Z., Tan Y., Li L., Zhou F., Hu S.A., Yan Q.W., Li L.H, & Pei G. (2024). Mechanism of Mulberry Leaves and Black Sesame in Alleviating Slow Transit Constipation Revealed by Multi-Omics Analysis. Molecules, 29(8), 1713.

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Extracellular Vesicle Protein Profiling

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EVs or tissue samples were lysed with radioimmunoprecipitation assay (RIPA) and proteinase inhibitor cocktail (Servicebio, Wuhan, China) at 4°C for 30 minutes. Then the lysates were centrifuged at 12,000 g for 20 minutes at 4°C. The protein concentration of EVs and Fallopian tube tissue samples were measured using a BCA Protein Assay Kit (Servicebio, Wuhan, China). 5μg of each sample was loaded for electrophoresis, using SDS/PAGE (10% gel), followed by transferring to PVDF membrane (Merck Millipore, Burlington, MA, USA). Then, the membrane was blocked using 5% skimmed milk for 1 h at room temperature, followed by washing with TBS for 15 min. Afterward, the membrane was incubated with the primary antibodies at 4 °C overnight. The primary antibodies used for immunostaining were TSG101 (1:1000; Abclonal, Woburn, MA, USA) and CD9 (1:1000; Abcam, Cambridge, UK), and the secondary antibodies were goat anti-rabbit labeled by horseradish peroxidase (1:2000; Servicebio, Wuhan, China). The membrane was incubated with the secondary antibodies for 1h at 37 °C and then was immersed in an electrochemiluminescence (ECL; Absin, Shanghai, China). The signals were detected using a Gene Gnome XRQ chemiluminescence imaging system (Syngene, Bengaluru, India).

Li Y., Cai L., Guo N., Liu C., Wang M., Zhu L., Li F., Jin L, & Sui C. (2023). Oviductal extracellular vesicles from women with endometriosis impair embryo development. Frontiers in Endocrinology, 14, 1171778.

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