Micro BCA Protein Assay Kit

Boster’s 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. Cited in 19 publication(s).

Product Info Summary

SKU AR1110
Pack size 1 kit
Applications Western blotting, protein expression assays, protein profiling and characterization, protein quantitation assays

List of Components

DescriptionQuantityVolumeContentsCatalog Number
Micro BCA Reagent A1100mL/AR1110-A
Micro BCA Reagent B1100mL/AR1110-B
Micro BCA Reagent C15mL/AR1110-C
Albumin (BSA) Standards120mL (2 mg/ml)containing bovine serum albumin (BSA) at 2mg/mL in 0.9% saline and 0.05% sodium azideAR1110-D

Overview

Form Supplied Micro BCA Reagent A, B, C: ready-to-use 1X solutions
BSA Standard: stock solution for preparing a series of standard protein dilutions
Assays per kit Tube procedure: 200 assays
Microplate procedure: 1300 assays
Storage Upon receipt store at room temperature.
It is stable at room temperature for one year.
Note If either Reagent MA or Reagent MB precipitates upon shipping in cold weather or during longterm storage, dissolve precipitates by gently warming and stirring solutions. Discard any reagent that shows discoloration or evidence of microbial contamination.
Assay Range Test Tube Procedure (linear working range of 0.5-20µg/mL)
Microplate Procedure (linear working range of 2-40µg/mL)
Equivalent Thermofisher (Product No. 23235),
Test Tube Procedure (linear working range of 0.5-20µg/mL)
Microplate Procedure (linear working range of 2-40µg/mL)
Millipore Sigma (Product No. QPBCA),
Test Tube Procedure (linear working range of 0.5-30µg/mL)
Microplate Procedure (linear working range of 1-20µg/mL)
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
Applications Western blotting, protein expression assays, protein profiling and characterization, protein quantitation assays
*Our Boster Guarantee covers the use of this product in the above tested applications.
Physical State of reagents mix Light blue liquid
Reagent Color/Absorbance Blue / A562 nm
Activity Active in alkaline medium
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.
The Micro BCA Protein Assay Kit provides an ultra-sensitive method to detect and quantify protein concentration. The kit has been optimized for determining total protein content in low protein concentration samples (0.5-20 μg/ml). The kit is sensitive to copper reductant and chelator instead of to both ionic and non-ionic detergents.
Cite This Product Micro BCA Protein Assay Kit (Boster Biological Technology, Pleasanton CA, USA, Catalog # AR1110)

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 Test tubes or miscroplates; Spectrophotometer; Microplate Reader
Assay Purpose Measure protein concentration in solution
Incubation conditions 60 min at 60°C

Assay Principle

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.
The Micro BCA Protein Assay Kit provides an ultra-sensitive method to detect and quantify protein concentration. The kit has been optimized for determining total protein content in low protein concentration samples (0.5-20 μg/ml). The kit is sensitive to copper reductant and chelator instead of to both ionic and non-ionic detergents.

Usage

Studying protein-protein interactions, Measuring column fractions after affinity chromatography, Assessing protein yields in whole cell lysates, High-throughput screening of fusion proteins.
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, Mitochondria Extraction Reagent, Membrane Protein Extraction Kit, Cytoplasmic and Nuclear Extraction Kit, Bacterial Cell Protein Extraction Reagent
For RIPA lysis buffer, dilution of 1:10 with distilled water is necessary to reduce the interference. It is recommended that same diluent should be used to make the BSA Standard solutions as that of the protein samples.

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.

Assay Protocol

Preparation of Standards and Working Reagent (required for both assay procedures)

A. Preparation of Albumin (BSA) Standards

Follow Table 1 to prepare a fresh set of standards. It is recommended that same diluent should be used to make the BSA Standard solutions as that of the protein samples.

Table1. Preparation of Diluted Albumin (BSA) Standards

Tube NumberVolume of Diluent (μl)Volume of BSA (μl)Final BSA Concentration (μg/ml)
A980 μl20 μl of 2 mg/ml Stock40 μg/ml
B400 μl400 μl of tube A 20 μg/ml
C400 μl400 μl of tube B10 μg/ml
D400 μl400 μl of tube C 5 μg/ml
E400 μl400 μl of tube D 2.5 μg/ml
F600 μl400 μl of tube E 1 μg/ml
G400 μl400 μl of tube F 0.5 μg/ml
H800 μl0 0 μg/ml =Blank

B. Preparation of BCA Working Reagent

1. Use the following formula to calculate the total volume of working reagent required: (well numbers for standard + well numbers for sample) × (times of repetition) × (volume of working reagent per well) = total volume of working reagent required

For example: For the standard Test Tube Procedure, each sample needs 3 dilutions and 2 replicates:

(8 standard wells + 3 sample wells) × (2 replicates) × (1 ml) = 22 ml working reagent required (round up to 25 ml)

Note: Each sample replicate requires 150 μl of BCA working reagent for each well in a microplate assay or 1 ml for a test tube procedure.

2. Mix Micro BCA Reagent A, Micro BCA Reagent B and Micro BCA Reagent C in the ratio of 25:24:1. i.e., mix 5 ml of Micro BCA Reagent A and 4.8 ml Micro BCA Reagent B with 0.2ml of Micro BCA Reagent C.

Note: When Micro BCA Reagent C is initially added into Micro BCA Reagent A and Micro BCA Reagent B, turbidity occurs that quickly disappears upon mixing to yield a clear-green solution.

Assay Procedure

• Test Tube Procedure

1. Add 1.0 ml of each standard and protein samples into separate labeled test tubes.

2. Add 1.0 ml of BCA working reagent to each tube and mix well.

3. Seal tubes and incubate at 60°C in a water bath for 1 hour.

4. Cool all tubes to room temperature (RT).

5. Set the wavelength of spectrophotometer at OD 562 nm. Calibrate the instrument to zero by using water. Subsequently, measure the absorbance of all samples within 10 minutes.
Note: Color development continues even after cooling to RT. However, the subsequent development at RT is too weak to produce significant error if all absorbance measurements are made within 10 minute.

6. Subtract OD562 of Blank from all readings.

7. Plot the BSA standard curve: OD562 (on Y axis) vs BSA Standard concentration (on X axis). Use the standard curve to determine the protein concentration of each unknown sample.

• Microplate Procedure

1. Add 150 μl of each standard and protein samples into separate microplate wells.

2. Add 150 μl of BCA working reagent to each well and mix thoroughly for 30 seconds.

3. Seal plates and incubate at 37°C for 2 hours or at 60°C for 1 hour.

4. Cool plate to room temperature (RT).

5. Measure the absorbance at 562 nm on a plate reader within 10 minutes.

6. Subtract OD562 of Blank from all readings.

7. Plot the BSA standard curve: OD562 (on Y axis) vs BSA Standard concentration (on X axis). Use the standard curve to determine the protein concentration of each unknown sample.

Product Images

AR1110 has been cited in 19 publications:

*The publications in this section are manually curated by our staff scientists. They may differ from Bioz's machine gathered results. Both are accurate. If you find a publication citing this product but is missing from this list, please let us know we will issue you a thank-you coupon.

Multi-functional zwitterionic coating for silicone-based biomedical devices

Mossa AH,Abdaem J,Cammisotto P,Campeau L.Deleterious impact of nerve growth factor precursor (proNGF) on bladder urothelial and smooth muscle cells.Cell Signal.2021 Jan 30:109936.doi:10.1016/j.cellsig.2021.109936.Epub ahead of print.PMID:33529756.
Species: Rat
AR1110 usage in article: APP:WB, SAMPLE:UROTHELIAL CELL, DILUTION:NA

The Novel Dual GLP-1/GIP Receptor Agonist DA-CH5 Is Superior to Single GLP-1 Receptor Agonists in the MPTP Model of Parkinson's Disease Lingyu Zhang J Parkinson's Dis. 2020;10(2):523-542. DOI: 10.3233/JPD-191768.

Autophagy protects bone marrow mesenchymal stem cells from palmitate%u2011induced apoptosis through the ROS%u2011JNK/p38 MAPK signaling pathways

Nanoparticle orientationally displayed antigen epitopes improve neutralizing antibody level in a model of porcine circovirus type 2

Expression of pannexin 1 and 2 in cortical lesions from intractable epilepsy patients with focal cortical dysplasia

Fas-associated factor 1 inhibits tumor growth by suppressing Helicobacter pylori-induced activation of NF-?B signaling in human gastric carcinoma

miR-20b Inhibits T Cell Proliferation and Activation via NFAT Signaling Pathway in Thymoma-Associated Myasthenia Gravis

Dihydrotestosterone modulates endothelial progenitor cell function via RhoA/ROCK pathway

Connective tissue growth factor stimulates the proliferation, migration and differentiation of lung fibroblasts during paraquat-induced pulmonary fibrosis

Mi B, Xiong W, Xu N, Guan H, Fang Z, Liao H, Zhang Y, Gao B, Xiao X, Fu J, Li F. Sci Rep. 2017 May 24;7(1):2328. doi: 10.1038/s41598-017-02491-9. Strontium-loaded titania nanotube arrays repress osteoclast differentiation through multiple signalli...

Tao R,, Wang ZF, Qiu W, He YF, Yan WQ, Sun WY, Li HJ. Exp Ther Med. 2017 Jun;13(6):2812-2818. doi: 10.3892/etm.2017.4294. Epub 2017 Apr 4. Role of S100A3 in human hepatocellular carcinoma and the anticancer effect of sodium cantharidinate

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 p...

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

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-Induc...

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.

Have you used Micro BCA Protein Assay Kit?

Submit a review and receive an Amazon gift card.

  • $25/€18/£15/$25CAN/¥75 Yuan/¥1250 Yen for a review with an image
  • $10/€7/£6/$10 CAD/¥70 Yuan/¥1110 Yen for a review without an image

Submit A Review

Be the first to review Micro BCA Protein Assay Kit

*The first user to submit a review for a product is eligible for Boster's Innovating Scientists Reward, which gives a free antibody. This is in addition to the gift card reward.

Have a question?

Find answers in Q&As, reviews.

Can't find your answer?

Submit your question

2 Customer Q&As for Micro BCA Protein Assay Kit

Question

What is the concentration and volume for the Micro BCA Reagent C: CuSO4 for AR1110?

Verified customer

Asked: 2019-06-18

Answer

The concentration and volume of the Micro BCA Reagent C: 4% CuSO4 for the Micro BCA Protein Assay Kit (AR1110) is 5ml.

Boster Scientific Support

Answered: 2019-06-19

Question

What are the components of the reagents for AR1110?

Verified customer

Asked: 2019-06-10

Answer

For the Micro BCA Protein Assay Kit (AR1110), the components of the reagents are; Micro BCA Reagent A: NaOH, Na2CO3, NaHCO3 Micro BCA Reagent B: BCA Micro BCA Reagent C:CuSO4

Boster Scientific Support

Answered: 2019-06-11

$80

Ask a question

Ships directly to USA and Canada. For other regions, visit our distributors page

Bulk Inquiry

Sample Inquiry

In stock
Order Product
AR1110
$80.00

Troubleshooting

troubleshooting-box-image

Download troubleshooting handbooks for IHC, Western blot and ELISA for FREE.

Download Free PDFs Now

Boster Guarantee

Guaranteed product quality

Guaranteed product quality

We promise all of our products perform as described in datasheets.