Superior colorimetric protein assay by minimizing interference from reducing agents.
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:
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):
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.
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.
Features of the Microplate BCA Protein Assay – RAC (Part No. 23252):
Protocol summary for Microplate BCA Protein Assay - Reducing Agent Compatible.
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.
