What is Bio-Rad reagent for protein assay?
The Bio-Rad Protein Assay is a dye-binding assay in which a differ- ential color change of a dye occurs in response to various concentrations of protein. 1 The absorbance maximum for an acidic solution of Coomassie® Brilliant Blue G-250 dye shifts from 465 nm to 595 nm when binding to protein occurs.
What is the Bio-Rad assay?
Description. The Bio-Rad protein assay is a simple colorimetric assay for measuring total protein concentration and is based on the Bradford dye-binding method (Bradford 1976). Using standard procedure, the assay is used with samples having protein concentrations between 200 and 1,400 µg/ml (20–140 µg total).
What is the purpose of the Bio-Rad dye reagent?
Use this Dye Reagent Concentrate to refill the Bio-Rad Protein Assay, a simple, accurate method for total protein measurement. This colorimetric assay based on the Bradford method is compatible with many common lab reagents.
Is RIPA buffer compatible with Bradford assay?
The RIPA buffer contains a mix of linear (SDS) and bulky (Igepal/Triton, deoxycholate) detergents, which makes it difficult to use in the Bradford assay.
How does Nanodrop measure protein concentration?
Using the absorbance at 280nm (A280), protein concentration (c) is calculated using the Beer-Lambert equation A280 = c * ε * b (ε is the wavelength-dependent protein extinction coefficient, b is the pathlength). Each pure protein has a unique extinction coefficient.
Does RIPA buffer interfere with BCA Assay?
Some protein kinases and other enzymes may be sensitive to the components of the RIPA Buffer, resulting in their decreased activity. In such cases, prepare a RIPA buffer that does not contain sodium deoxycholate and SDS. RIPA Buffer is compatible with the Thermo Scientific™ Pierce™ BCA Protein Assay Kit (Cat. No.
Does RIPA buffer interfere with BCA assay?
What is BSA buffer?
Bovine serum albumin (BSA) blocking buffer is ideal for saturating excess protein-binding sites on membranes and microplates for Western blotting and ELISA applications, respectively. Typically, 1-3% BSA is sufficient for most applications.
What does BSA block?
Abstract. BSA blocking is a routine practice among clinicians and researchers working on immunoassays throughout the world. The primary role of BSA is to prevent the non-specific binding by blocking the leftover spaces over solid surface after immobilization of a capture biomolecule.
What is the working reagent in BCA assay?
Principle. BCA serves the purpose of the Folin reagent in the Lowry assay, namely to react with complexes between copper ions and peptide bonds to produce a purple end product.
Does EDTA interfere with BCA assay?
EDTA and reducing agents, such as DTT, BME, TCEP and other disulfide reducing agents, interfere with the BCA assay. The concentration of proteins can be also overestimated if proteins form soluble aggregates in solutions.
Why is BSA used in BCA assay?
The intensity of the color formed by these assays is measured by absorbance photometry at 595 nm and 562 nm for the Bradford and BCA assays, respectively. Typically, standard solutions of bovine serum albumin (BSA) are used to produce a calibration curve of absorbance versus mass concentration.
What does assay buffer do?
Assay buffers are mixtures of buffers, salts, detergents, protein, preservatives, and coloring agents. These buffers are used in ELISA assays for dilution of standards, samples, and reagents, and for blocking of non-specific interactions.