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SKU AR1110
Pack size 1 kit
Applications Western blotting, protein expression assays, protein profiling and characterization, protein quantitation assays
Price: $80.00
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Overview

Product Name Micro BCA Protein Assay Kit
Synonyms Micro Bicinchoninic acid protein assay kit; Micro BCA Protein Quantification Kit, Micro BCA Protein Quantitation Kit, Micro BCA Protein Estimation Kit, Micro Bicinchoninic acid Protein Quantification Kit, Micro Bicinchoninic acid Protein Quantitation Kit, Micro Bicinchoninic acid Protein Estimation Kit, Micro BCA kit for protein determination, Micro Bicinchoninic acid kit for protein determination, Smith Protein Assay kit
Description Micro 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 of dilute protein solutions by measuring absorbance at 562 nm and comparing to a protein standard absorbance vs. concentration curve.
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 0.5 – 20 μg/ml
Incubation conditions 60 min at 60°C
Kit components BCA Reagent A: 100 ml
BCA Reagent B: 100 ml
BCA Reagent C: 5 ml
Albumin (BSA) Standards: 20 ml (2 mg/ml)

Properties

Form Supplied BCA Reagent A, B, C: ready-to-use 1X solutions
BSA Standard: stock solution for preparing a series of standard protein dilutions
Physical State of reagents mix 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: 100 assays
Microplate procedure: 300 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 (e.g. copper reductants): 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 Micro BCA Assay uses concentrated solutions and extended incubation times for the detection of dilute protein samples.
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 protein present. 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.

Micro BCA Protein Assay Kit Images

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Micro BCA Protein Assay Kit
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Publications

Wang, X., An, F., Wang, S., An, Z., & Wang, S. (2017). Orientin Attenuates Cerebral Ischemia/Reperfusion Injury in Rat Model through the AQP-4 and TLR4/NF-κB/TNF-α Signaling Pathway. Journal of Stroke and Cerebrovascular Diseases. Advance online publication. doi: 10.1016/j.jstrokecerebrovasdis.2017.05.002
Yan Ding, Wang Liao, Xiao-Jie He, Wei Xiang First published: November 2016 DOI: 10.1111/jcmm.13037 Effects of 1,25(OH)2D3 and vitamin D receptor on peripheral CD4+/CD8+ double-positive T lymphocytes in a mouse model of systemic lupus erythematosus
J Liu, X Nie, Y Shao, W Su, H Ma, X Xu - Cellular Physiology and Biochemistry, 2016 Bleomycin Suppresses the Proliferation and the Mobility of Human Gastric Cancer Cells Through the Smad Signaling Pathway
Hong-Yan Zhanga, Feng Liangb Fei Wanga, Jian-Wei Zhanga, Li Wanga,Xi-Gang Kanga, Jie Wanga, Qi-Liu Duana Cell Physiol Biochem 2016;40:807-817 In Vitro Effects of HAS-2 Gene Silencing on the Proliferation and Apoptosis of the MCF-7 Human Breast Cancer Cell Line
Luo J. Y. · Fu D. · Wu Y. Q. · Gao Y. Cell Physiol Biochem 2016;40:527-537 (DOI:10.1159/000452566) Inhibition of the JAK2/STAT3/SOSC1 Signaling Pathway Improves Secretion Function of Vascular Endothelial Cells in a Rat Model of Pregnancy-Induced Hypertension
Li X, Luan S, Yuan S, Song L, Zhao J, Ma J, Shi H, Yang H, Jin J, Yin J. Colloids Surf B Biointerfaces. 2013 Dec 1;112:146-54. Doi: 10.1016/J.Colsurfb.2013.07.048. Epub 2013 Aug 3. Surface Functionalization Of Styrenic Block Copolymer Elastomeric ...
Liu Y, Yu Y, Chu H, Bing D, Wang S, Zhou L, Chen J, Chen Q, Pan C, Sun Y, Cui Y. Neurosci Lett. 2015 Feb 19;588:72-7. Doi: 10.1016/J.Neulet.2014.12.060. Epub 2014 Dec 31. 17-Dmag Induces Hsp70 And Protects The Auditory Hair Cells From Kanamycin Ot...
Sun H, Liu K, Liu W, Wang W, Guo C, Tang B, Gu J, Zhang J, Li H, Mao X, Zou Q, Zeng H. Int J Nanomedicine. 2012;7:5529-43. Doi: 10.2147/Ijn.S36071. Epub 2012 Oct 26. Development And Characterization Of A Novel Nanoemulsion Drug-Delivery System For...
Zhao J, Song L, Shi Q, Luan S, Yin J. Acs Appl Mater Interfaces. 2013 Jun 12;5(11):5260-8. Doi: 10.1021/Am401098U. Epub 2013 May 20. Antibacterial And Hemocompatibility Switchable Polypropylene Nonwoven Fabric Membrane Surface.