Why is BSA used in protein estimation?
BSA is used because of its ability to increase signal in assays, its lack of effect in many biochemical reactions, and its low cost, since large quantities of it can be readily purified from bovine blood, a byproduct of the cattle industry.
Why is BSA used in BCA assay?
Typically, standard solutions of bovine serum albumin (BSA) are used to produce a calibration curve of absorbance versus mass concentration. Assuming the analyte-proteins react in the same manner as the BSA standard, the unknown concentration can be determined.
Why is BSA not used in Bradford assay?
units. Perhaps you were told this because BSA is not a suitable standard for some purified proteins due to a substantial difference is amino acid compositions. For example, the slopes of Bradford assay standard curved for BSA and immunoglobulin differ significantly.
What does BCA assay measure?
Bicinchoninic acid (BCA) protein assays. The BCA Protein Assay was introduced by Smith, et al., in 1985. Since then it has become the most popular method for colorimetric detection and quantitation of total protein.
What does BSA assay stand for?
Bovine serum albumin
Bovine serum albumin (BSA) is a globular protein that is often used as a protein concentration standard in lab experiments as well as in numerous other biochemical applications. It has been prevalent in the use of protein assays since the Bradford Protein Assay was first introduced in 1976.
Why is BSA used in biuret test?
An aqueous solution of bovine serum albumin (BSA) is commonly used as a standard in testing because of its stability in testing, low cost, and is readily available as a byproduct from bovine blood.
Why BSA is used as a standard?
BSA is used because of its stability to increase signal in assays, its lack of effect in many biochemical reactions, and its low cost, since large quantities of it can be readily purified from bovine blood, a byproduct of the cattle industry.
Is BSA a good standard for Bradford assay?
Prepare a standard curve of absorbance versus micrograms protein and determine amounts from the curve. *Note: The ubiquitously used BSA is a very poor standard for the Bradford assay.
Is the BSA a good standard to use in the Bradford assay?
Therefore, the BSA standard would be an appropriate standard if the sample contains primarily albumin, or if the protein being assayed gives similar response to the dye. For a color response that is typical of many proteins, the gamma-globulin standard is appropriate.
How does BCA assay calculate protein concentration?
You can use this formula to calculate the amount of protein in your sample: [ (OD of the test sample) / (OD of the standard sample) ] X concentration of the standard sample. = 4.0 mg/mL protein (in test sample).
What is the detection range of proteins by BCA assay?
20 to 2,000 μg/mL
The standard Pierce BCA Protein Assay detects protein concentrations from 20 to 2,000 μg/mL using a two-component system: Reagent A, a carbonate buffer containing BCA reagent, and Reagent B, a cupric sulfate solution, which are combined to make an apple green–colored working solution that turns purple after 30 minutes …
Why is protein assay important?
The measurement of protein concentration in an aqueous sample is an important assay in biochemistry research and development labs for applications ranging from enzymatic studies to providing data for biopharmaceutical lot release.
How does the Bradford protein assay work?
Bradford protein assay. The Bradford protein assay was developed by Marion M. Bradford in 1976. It is a quick and accurate spectroscopic analytical procedure used to measure the concentration of protein in a solution. The reaction is dependent on the amino acid composition of the measured protein.
What is a protein assay?
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).