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SKU AR0146
Pack size 1 kit
Applications Western blotting, protein expression assays, protein profiling and characterization, protein quantitation assays

Overview

Product Name BCA Protein Assay Kit
Synonyms Bicinchoninic acid protein assay kit; BCA Protein Quantification Kit, BCA Protein Quantitation Kit, BCA Protein Estimation Kit, Bicinchoninic acid Protein Quantification Kit, Bicinchoninic acid Protein Quantitation Kit, Bicinchoninic acid Protein Estimation Kit, BCA kit for protein determination, Bicinchoninic acid kit for protein determination, Smith Protein Assay kit
Description BCA Protein Assay Kit is a ready-to-use detergent-compatible Western blot related total protein analysis reagent used for the quick determination of total protein concentration by measuring absorbance at 562 nm and comparing to a protein standard absorption vs. concentration curve, according to Smith. The protein quantification process can be finished in 45 minutes.
Application Western blotting, protein expression assays, protein profiling and characterization, protein quantitation assays
Pack Size 1 kit
Reagent Type Protein assay kit; Western Blotting related
Usage Studying protein-protein interactions, Measuring column fractions after affinity chromatography, Assessing protein yields in whole cell lysates, High-throughput screening of fusion proteins

Assay Information

Assay Type Protein Quantitative Measurement
Sample Type Serum, Plasma, Cell culture extracts, Tissue Extracts
Technique Solution-based detection of spectral absorbance at 562 nm; Smith Protein assay
Detection Method Colorimetric/Spectroscopic
Equipment needed Тest tubes or miscroplates; Spectrophotometer; Microplate Reader
Assay Purpose Measure protein concentration in solution
Assay Range 20 - 2000 μg/ml
Incubation Requirements 30 min at 37°C
Kit components BCA Reagent A: 100 ml
BCA Reagent B: 5 ml
Albumin (BSA) Standards: 20 ml (2 mg/ml)

Properties

Form Supplied BCA Reagent A and B: ready-to-use 1X solutions
BSA Standard: stock solution for preparing a series of standard protein dilutions
Physical State of A+B Reagent mixture Light blue liquid
Recommended working concentration See protocol in Datasheet
Reagent Color/Absorbance Blue / A562 nm
Activity Active in alkaline medium
Assays per kit Tube procedure: 50 assays
Microplate procedure: 500 assays
Compatibility with reagents Compatible with typical concentrations of most ionic and nonionic detergents
Incompatible with chelating agents, strong acids or bases, and reducing agents: interfere with the reduction and chelating reactions of the assay mechanism
Storage Store at room temperature for one year.
Precautions FOR RESEARCH USE ONLY. NOT FOR DIAGNOSTIC AND CLINICAL USE

Usage and Handling

To be used with: Lysates or homogenates, prepared with non-reducing lysis and extraction buffers: Mammal Tissue Protein Extraction Reagent, Mammal Cell Protein Extraction Reagent, RIPA Lysis Buffer, Mitochondria Extraction Reagent, Membrane Protein Extraction Kit, Cytoplasmic and Nuclear Extraction Kit, Bacterial Cell Protein Extraction Reagent

Background

The BCA Protein Assay is a detergent - compatible formulation based on bicinchoninic acid (BCA) for the colorimetric detection and quantization of total protein. This method combines the reduction of Cu 2+ to Cu 1+ by protein in an alkaline medium (Biuret reaction) with the highly sensitive and selective colorimetric detection of the cuprous cation (Cu 1+) by bicinchoninic acid, forming a purple-colored BCA/copper complex with absorbance maximum at 562 nm. Measurement of A562 is used to determine the concentration of protein in solutions. Protein concentrations are estimated by reference to absorbances obtained for a series of standard protein dilutions (e.g. of BSA), which are assayed alongside the unknown sample. The BCA assay involves two consecutive steps of color development, where the final product indirectly depends on the protein concentration in the solution. The first step is the chelation of copper with protein in an alkaline environment, followed by reduction of the Cu2+ (cupric) to Cu1+ (cuprous) cations forming a light blue complex. It has been shown that cysteine, cystine, tryptophan, tyrosine, and the peptide bond are able to reduce Cu2+ to Cu1+. The amount of reduction is proportional to the amount of protein present (the number of peptide bonds). In the second step of the color development reaction, bicinchoninic acid (BCA), a highly specific chromogenic reagent for Cu1+, reacts with the reduced cuprous cation that was formed in step one, producing a purple complex with strong absorbance at 562 nm (Smith 1985, Wiechelman 1988). The purple-colored reaction product results from the chelation of two molecules of BCA with one cuprous ion. The intensity of the BCA/copper purple complex is proportional to the amount of protein in the sample. Thus measuring the intensity of the absorbance at 562 nm is proportional to protein concentration. The BCA protein assay demonstrates higher tolerances towards common interfering substances, such as nonionic detergents and buffer salts, than the Lowry technique.

BCA Protein Assay Kit Images

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BCA Protein Assay Kit
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Publications

Li Q, Ding L, Jing N, Liu C, Yang Z, Chen F, Hou L, Wang J. Oncol Lett. 2018 Feb;15(2):1963-1972. doi: 10.3892/ol.2017.7471. Epub 2017 Nov 22. Folate deficiency and aberrant DNA methylation and expression of FHIT gene were associated with cervical pathogenesis
An J, Zhang Z, Liu Z, Wang R, Hui D, Jin Y. BMC Cancer. 2017 Dec 6;17(1):828. doi: 10.1186/s12885-017-3839-7. Overexpression of Cullin7 is associated with hepatocellular carcinoma progression and pathogenesis
Du Y, Li J, Xu T, Zhou DD, Zhang L, Wang X. Oncotarget. 2017 Jun 22;8(37):61510-61527. doi: 10.18632/oncotarget.18604. eCollection 2017 Sep 22. MicroRNA-145 induces apoptosis of glioma cells by targeting BNIP3 and Notch signaling
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Xu C, Li X, Guo P, Wang J. Med Sci Monit. 2017 Oct 14;23:4909-4917. Hypoxia-Induced Activation of JAK/STAT3 Signaling Pathway Promotes Trophoblast Cell Viability and Angiogenesis in Preeclampsia
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Zhang L, Xu C, Hu W, Wu P, Qin C, Zhang J. Int J Mol Med. 2018 Mar;41(3):1293-1304. doi: 10.3892/ijmm.2017.3327. Epub 2017 Dec 18. Anti-inflammatory effects of Lefty-1 in renal tubulointerstitial inflammation via regulation of the NF-
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Liu LJ, Yu JJ, Xu XL. Am J Transl Res. 2017 Jul 15;9(7):3499-3513. eCollection 2017. MicroRNA-93 inhibits apoptosis and promotes proliferation, invasion and migration of renal cell carcinoma ACHN cells via the TGF-β/Smad signaling pathway by targeting RUNX3
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Bao W, Luo Y, Wang D, Li J, Wu X, Mei W. J Cell Biochem. 2018 Jan;119(1):850-860. doi: 10.1002/jcb.26249. Epub 2017 Aug 30. Sodium salicylate modulates inflammatory responses through AMP
Fang X, Han Q, Li S, Zhao Y, Luo A. Am J Transl Res. 2017 Sep 15;9(9):4288-4299. eCollection 2017. Chikusetsu saponin IVa attenuates isoflurane-induced neurotoxicity and cognitive deficits via SIRT1/ERK1/2 in developmental rats
Dai B, Zhu F, Chen Y, Zhou R, Wang Z, Xie Y, Wu X, Zu S, Li G, Ge J0, Chen F. Int J Mol Sci. 2017 Oct 11;18(10). pii: E2125. doi: 10.3390/ijms18102125. ASIC1a Promotes Acid-Induced Autophagy in Rat Articular Chondrocytes through the AMPK/FoxO3a Pathway
Liu S, Li X, Wu Y, Duan R, Zhang J, Du F, Zhang Q, Li Y, Li N. PLoS One. 2017 Dec 14;12(12):e0189722. doi: 10.1371/journal.pone.0189722. eCollection 2017. Effects of vaspin on pancreatic β cell secretion via PI3K/Akt and NF-κB signaling pathways
Kong LY, Xue M, Zhang QC, Su CF. Oncotarget. 2017 Feb 28;8(9):15507-15519. doi: 10.18632/oncotarget.14662. In vivo and in vitro effects of microRNA-27a on proliferation, migration and invasion of breast cancer cells through targeting of SFRP1 gene via Wnt/β-catenin signaling pathway
Liu B, Liu C, Cong W, Li N, Zhou N, Tang Y, Wei C, Bai H, Zhang Y, Xiao J. Skelet Muscle. 2017 May 11;7(1):8. doi: 10.1186/s13395-017-0126-x. Retinoid acid-induced microRNA-31-5p suppresses myogenic proliferation and differentiation by targeting CamkII
Wang R, Shen Z, Yang L, Yin M, Zheng W, Wu B, Liu T, Song H. Int J Mol Med. 2017 Nov;40(5):1537-1548. doi: 10.3892/ijmm.2017.3121. Epub 2017 Sep 6. Protective effects of heme oxygenase-1-transduced bone marrow-derived mesenchymal stem cells on reduced-size liver transplantation: Role of autophagy regulated by the ERK/mTOR signaling pathway
Huang L, Zhang K, Guo Y, Huang F, Yang K, Chen L, Huang K, Zhang F, Long Q, Yang Q. Sci Rep. 2017 Sep 20;7(1):11989. doi: 10.1038/s41598-017-12095-y. Honokiol protects against doxorubicin cardiotoxicity via improving mitochondrial function in mouse hearts
Feng XE, Liang TG, Gao J, Kong P, Ge R, Li QS,. Oxid Med Cell Longev. 2017;2017:7028478. doi: 10.1155/2017/7028478. Epub 2017 Jun 14. Heme Oxygenase-1, a Key Enzyme for the Cytoprotective Actions of Halophenols by Upregulating Nrf2 Expression via Activating Erk1/2 and PI3K/Akt in EA.hy926 Cells
Zhou ZQ, Wang ZK, Zhang L, Ren YQ, Ma ZW, Zhao N, Sun FY. Biosci Rep. 2017 Jul 27;37(4). pii: BSR20170021. doi: 10.1042/BSR20170021. Print 2017 Aug 31. Role of ESAT-6 in renal injury by regulating microRNA-155 expression via TLR4/MyD88 signaling pathway in mice with Mycobacterium tuberculosis infection
Tang S, Pan H, Wei W, Yang H, Liu J, Yang R. Oncotarget. 2017 Oct 19;8(62):105155-105169. doi: 10.18632/oncotarget.21927. eCollection 2017 Dec 1. GOLPH3: a novel biomarker that correlates with poor survival and resistance to chemotherapy in breast cancer
Kuang X, Zhou HJ, Thorne AH, Chen XN, Li LJ, Du JR. Front Aging Neurosci. 2017 Nov 2;9:353. doi: 10.3389/fnagi.2017.00353. eCollection 2017. Neuroprotective Effect of Ligustilide through Induction of α-Secretase Processing of Both APP and Klotho in a Mouse Model of Alzheimer’s Disease
Li X, Wei K, Hu R, Zhang B, Li L, Wan L, Zhang C, Yao W. Front Cell Neurosci. 2017 Nov 23;11:368. doi: 10.3389/fncel.2017.00368. eCollection 2017. Upregulation of Cdh1 Attenuates Isoflurane-Induced Neuronal Apoptosis and Long-Term Cognitive Impairments in Developing Rats
Wu YX, Sun Y, Ye YP, Zhang P, Guo JC, Huang JM, Jing XZ, Xiang W, Yu SY, Guo FJ. Mol Med Rep. 2017 Dec;16(6):8200-8208. doi: 10.3892/mmr.2017.7648. Epub 2017 Sep 28. Iguratimod prevents ovariectomy-induced bone loss and suppresses osteoclastogenesis via inhibition of peroxisome proliferator-activated receptor-γ
Huang JM, Bao Y, Xiang W, Jing XZ, Guo JC, Yao XD, Wang R, Guo FJ. Evid Based Complement Alternat Med. 2017;2017:8085325. doi: 10.1155/2017/8085325. Epub 2017 Dec 27. Icariin Regulates the Bidirectional Differentiation of Bone Marrow Mesenchymal Stem Cells through Canonical Wnt Signaling Pathway
Ding Y, Liao W, He XJ, Xiang W. J Cell Mol Med. 2017 May;21(5):975-985. doi: 10.1111/jcmm.13037. Epub 2017 Jan 7. Effects of 1,25(OH)2D3 and vitamin D receptor on peripheral CD4+/CD8+ double
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Li YL, Li YF, Li HF, Lv HQ, Sun DZ. Biosci Rep. 2017 Jul 12;37(4). pii: BSR20170144. doi: 10.1042/BSR20170144. Print 2017 Aug 31. Role of SDF-1α/CXCR4 signaling pathway in clinicopathological features and prognosis of patients with nasopharyngeal carcinoma
Luo HT, Liang CX, Luo RC, Gu WG. Biosci Rep. 2017 Nov 6;37(6). pii: BSR20171086. doi: 10.1042/BSR20171086. Print 2017 Dec 22. Identification of relevant prognostic values of cytokeratin 20 and cytokeratin 7 expressions in lung cancer
Li YF, Yuan YY, Zhang YM, Zhao N, Zhang Q, Meng FX, Gao RP, Yu BF, Zhang YH, Guo R, Wang HL, Xie J, Xu J, Qin Q, Dong XS. Mol Med Rep. 2017 Jul;16(1):764-772. doi: 10.3892/mmr.2017.6657. Epub 2017 May 31. HSVtk/GCV system on hepatoma carcinoma cells: Construction of the plasmid pcDNA3.1-pAFP-TK and targeted killing effect
Luo J, Liu YH, Luo W, Luo Z, Liu CT. Sci Rep. 2017 Jan 4;7:39905. doi: 10.1038/srep39905. β2-adrenoreceptor Inverse Agonist Down-regulates Muscarine Cholinergic Subtype-3 Receptor and Its Downstream Signal Pathways in Airway Smooth Muscle Cells in vitro
Bailin N, Nan C, Peizhi L, Kun H, Xiwen Z, Guosheng R, Jianping G, Wenfeng Z. Oncotarget. 2017 Aug 7;8(44):76783-76796. doi: 10.18632/oncotarget.20011. eCollection 2017 Sep 29. Changes of Foxo3a in PBMCs and its associations with stress hyperglycemia in acute obstructive suppurative cholangitis patients
Li K, Gesang L, He C. Exp Ther Med. 2017 Oct;14(4):3780-3787. doi: 10.3892/etm.2017.4996. Epub 2017 Aug 22. Mechanism of apoptosis involved in gastric mucosal lesions in Tibetans with high-altitude polycythemia
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Wang G, Hu Z, Fu Q, Song X, Cui Q, Jia R, Zou Y, He C, Li L, Yin Z. Sci Rep. 2017 Mar 21;7:45006. doi: 10.1038/srep45006. Resveratrol mitigates lipopolysaccharide-mediated acute inflammation in rats by inhibiting the TLR4/NF-
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He XJ, Bian EB, Ma CC, Wang C, Wang HL, Zhao B. Oncol Lett. 2018 Mar;15(3):3977-3984. doi: 10.3892/ol.2018.7776. Epub 2018 Jan 12. Long non-coding RNA SPRY4-IT1 promotes the proliferation and invasion of U251 cells through upregulation of SKA2
Zhang Y, Yang B, Zhao J, Li X, Zhang L, Zhai Z. Med Sci Monit. 2018 Jan 15;24:294-304. Proteasome Inhibitor Carbobenzoxy-L-Leucyl-L-Leucyl-L-Leucinal (MG132) Enhances Therapeutic Effect of Paclitaxel on Breast Cancer by Inhibiting Nuclear Factor (NF)-
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Gao J, Luo A, Yan J, Fang X, Tang X, Zhao Y, Li S. Am J Transl Res. 2018 Feb 15;10(2):432-443. eCollection 2018. Mdivi-1 pretreatment mitigates isoflurane-induced cognitive deficits in developmental rats.
Kai Xu, Qingsheng Niu, Huiting Zhao, Yali Du, Lina Guo, and Yusuo Jiang. Journal of Insect Science. Sequencing and Expression Characterization of Antifreeze Protein Maxi-Like in Apis cerana cerana.
Junfa Yang, Qingxue Liu, Shiyang Cao, Tao Xu, Xiaofeng Li, Dandan Zhou, Linxin Pan, Changyao Li, Cheng Huang, Xiaoming Meng, Lei Zhang, and Xiao Wang. Front Pharmacol. 2018 Jan 12;8:980. doi: 10.3389/fphar.2017.00980. eCollection 2017. MicroRNA-145 Increases the Apoptosis of Activated Hepatic Stellate Cells Induced by TRAIL through NF-κB Signaling Pathway.
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Ling, Q., Yu, X., Wang, T., Wang, S.G., Ye, Z.Q., & Liu, J.H. (2017). Roles of the Exogenous H2S-Mediated SR-A Signaling Pathway in Renal Ischemia/ Reperfusion Injury in Regulating Endoplasmic Reticulum Stress-Induced Autophagy in a Rat Model. Cellular Physiology and Biochemistry, 41(6), 2461-2474. doi: 10.1159/000475915.
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Customer Q&As

Q: What is the correct storage temperature of this kit? On the manual it says room temperature, but in the manual that we received in the past it said 4 degrees.
A: We have modified the storage temperature of this kit as room temperature recently, but it can be stored at 4 degree as well. The BSA solution contains sodium azide, it is available at room temperature.