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BCA Protein Assay - Reducing Agent Compatible


Superior colorimetric protein assay by minimizing interference from reducing agents.

BCA Protein Assay Kit, Reducing Agent Compatible (BCA-RAC)

This reducing agent-compatible version of our popular Thermo Scientific Pierce BCA Protein Assay enables one to measure protein concentration in samples that contain thiol-reductants dithiothreitol (DTT) and 2-mercaptoethanol (BME).

The BCA Protein Assay Kit – Reducing Agent Compatible (BCA-RAC) provides all of the advantages of the original BCA Assay, plus compatibility with disulfide reducing agents at concentrations routinely used in protein sample buffers. This special adaptation of the popular Pierce BCA Protein Assay method enables accurate protein concentration measurement for samples containing DTT, 2-ME or TCEP. Available in tube and microplate formats, these reducing agent compatible (RAC) BCA Kits extend the already broad reagent compatibility of the BCA Protein Assay protocol to include nearly all types of components commonly present in protein research samples.

Highlights:

  • Compatible – assay samples that contain up to 5mM DTT, 35mM BME, or 10mM TCEP
  • BCA technology – only a slight modification of the standard BCA Protein Assay protocol (15-minute incubation with Compatibility Reagent); no precipitation steps required
  • Small samples – requires only 25µL (standard kit) or less than 10µL (microplate kit) of sample
  • Colorimetric – measure with a standard spectrophotometer or plate reader (562nm)
  • Excellent uniformity – exhibits less protein-to-protein variation than dye-binding methods
  • High linearity – linear working range for BSA equals 125 to 2000µg/mL

Product Details:

Video of the Pierce BCA-RAC Protein Assay Kit in action!

 

Comparison of the Reducing Agent-Compatible BCA Assays.
  Microplate BCA-RAC
(Part No. 23252)
Tube BCA-RAC
(Part No. 23250)
Sample Requirement Nine microliters (9µL) Twenty-five microliters (25µL)
Validated Microplates Includes 20 microplates Not included
Compatibility Reagent Supplied as 48 x 10mg microtubes Supplied as 10 x 20mg vials

Features of BCA Protein Assay – RAC (Part No. 23250):

Effect of DTT on the standard BCA Protein Assay Compatibility of DTT on the Protein Assay Reducing Agent Compatible
A. Standard BCA Protein Assay B. BCA-RAC Protein Assay
 

Comparison of the effect of reducing agents on Standard BCA and BCA-RAC protein assays. Reducing agents (>1mM) interfere with the standard BCA Protein Assay by artificially increasing the color intensity of all samples regardless of protein concentration (left panel). The color intensity produced by the BCA Protein Assay – Reducing Agent Compatible (BCA-RAC) is not artificially increased by the presence of reducing agents (right panel). The diagram illustrates the effect of samples containing 5mM DTT and BSA standards at the following concentrations (µg/mL) for tubes 1-8, respectively: 0, 125, 250, 500, 750, 1000, 1500, 2000.

 
Protocol summary of the Thermo Scientific BCA Protein Assay Reducing Agent Compatible (BCA-RAC)

 

Color response curves generated using BSA after treatment with Compatibility Reagent in the presence and absence of 5mM DTT

Color response curves generated using BSA after treatment with Compatibility Reagent in the presence and absence of 10 mM TCEP

BCA-RAC Protein Assay produces a linear standard curve in the presence of reducing agents. Color response curves for BSA after treatment with Compatibility Reagent in the presence and absence of 5mM DTT, 35mM 2-mercaptoethanol (2ME or BME) and 10mM TCEP.

Color response curves generated using BSA after treatment with Compatibility Reagent in the presence and absence of 35mM 2-mercaptoethanol

Features of the Microplate BCA Protein Assay – RAC (Part No. 23252):

Microplate BCA Protein Assay - Reducing Agent Compatible Protocol
Microplate BCA Protein Assay - Reducing Agent Compatible Protocol
Protocol summary for Microplate BCA Protein Assay - Reducing Agent Compatible.

 

Color response curves generated using BSA after treatment with Compatibility Reagent in the presence and absence of 5mM DTT Microplate BCA-RAC Assay produces linear standard curves in the presence of reducing agents. Color response curves generated using BSA after treatment with Compatibility Reagent in the presence and absence of 5mM DTT, 25mM 2-mercaptoethanol (2ME or BME) and 10mM TCEP.
Color response curves generated using BSA after treatment with Compatibility Reagent in the presence and absence of 25 mM 2-Mercaptoethanol
Color response curves generated using BSA after treatment with Compatibility Reagent in the presence and absence of 10 mM TCEP

 

Related Resources:

BCA Protein Assay - RAC FAQ
Protein Assay Selection Guide
Protein Assay Compatibility Table

Protein Methods Library:

Overview of Protein Assays
Chemistry of Protein Assays
Protein Assay Data Analysis

Compatible Instrumentation:

BioMate 3 UV-Vis Spectrophotometer
NanoDrop 2000c UV/Vis Spectrophotometer
NanoDrop 8000 UV/Vis 8 Sample Spectrophotometer

Related Products:

BSA Assay StandardsBGG Assay Standards
BCA, Micro BCA, and Other Assay Reagents
Microplates for Protein Assays


Ordering Information
 
Product # Description Pkg. Size Instructions MSDS CofA Price
23250 BCA Protein Assay Kit - Reducing Agent Compatible
Formulation: Multi-component kit
Sufficient For: 250 tube assays with 25µL samples

Kit Contents:
BCA Reagent A, 250mL
BCA Reagent B, 25mL
Compatibility Reagent, 10 x 20mg
Reconstitution Buffer, 15mL
Albumin Standard Ampules, 2mg/mL, 10 x 1mL
275mL kit Product Instructions for product #23250 BCA Protein Assay Kit - Reducing Agent Compatible MSDS for product #23250 BCA Protein Assay Kit - Reducing Agent Compatible Certificate of Analysis for product #23250 BCA Protein Assay Kit - Reducing Agent Compatible $218.00

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23252 Microplate BCA Protein Assay Kit - Reducing Agent Compatible
Formulation: Multi-component kit
Sufficient For: 980 microplate assays with 9µL samples

Kit Contents:
BCA Reagent A, 250mL
BCA Reagent B, 25mL
Compatibility Reagent, 48 x 10mg
Reconstitution Buffer, 15mL
Albumin Standard Ampules, 2mg/mL, 10 x 1mL
96-Well Microplates, 20 plates
275mL kit Product Instructions for product #23252 Microplate BCA Protein Assay Kit -  Reducing Agent Compatible MSDS for product #23252 Microplate BCA Protein Assay Kit -  Reducing Agent Compatible Certificate of Analysis for product #23252 Microplate BCA Protein Assay Kit -  Reducing Agent Compatible $218.00

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15045 96-Well Plates for BCA-RAC Assay
Formulation: Polystyrene; 96-well; round-bottom
Sufficient For: Use with the BCA-RAC Assay (Part No. 23252)
5 plates $35.00

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Instructions | MSDS | CofA
Product Instructions | MSDS | CofA  

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