What are the steps of BCA protein assay?
What are the steps of BCA protein assay?
Assay
- Prepare samples containing 0.2 to 50 micrograms protein in microliters.
- Add 1 ml SWR to each 20 microliters sample and mix. Incubate 30 min. at 60 degrees C.
- Cool the samples and read at 562 nm. Color will be stable for at least one hour.
Why is the BCA method used for the protein quantitative analysis?
The BCA assay has a lot of advantages. Compared to other methods, the BCA assay is one of the most sensitive (it can detect proteins at concentrations as low as 5 ug/mL). It has less variability than others (i.e., Bradford assay), and it can be used to measure a wide range of protein concentration.
How does BCA test work?
The BCA assay primarily relies on two reactions. First, the peptide bonds in protein reduce Cu2+ ions from the copper(II) sulfate to Cu1+ (a temperature dependent reaction). The amount of Cu2+ reduced is proportional to the amount of protein present in the solution.
What is BCA assay test?
The Pierce BCA assay can be used to assess yields in whole cell lysates, affinity-column fractions, purified proteins samples, as well as to monitor protein contamination in industrial applications.
How long does a BCA assay take?
Choose the right BCA protein assay for your sample
| Rapid Gold BCA Protein Assay | Micro BCA Protein Assay | |
|---|---|---|
| Incubation time and temperature | 5 min at RT | 60 min at 60°C |
| Total assay time | 50 min | 130 min |
| Absorbance | 480 nm | 562 nm |
| Compatible reagents | Detergents | Detergents |
What is BCA reagent?
BCA Protein Assay Reagent A is a component of the Pierce BCA Protein Assay Kit, a two-component, high-precision, detergent-compatible assay that is used for total protein concentration determination compared to a protein standard.
Which method is best for protein estimation?
The simplest and most direct assay method for protein concentration determination in solution is to measure the absorbance at 280 nm (UV range). Amino acids containing aromatic side chains (i.e., tyrosine, tryptophan and phenylalanine) exhibit strong UV-light absorption.
What is the Pierce BCA protein assay based on?
bicinchoninic acid
The Thermo Scientific™ Pierce™ BCA Protein Assay Kit is a detergent-compatible formulation based on bicinchoninic acid (BCA) for the colorimetric detection and quantitation of total protein.
What is BCA chemical?
Bicinchoninic acid is most commonly employed in the bicinchoninic acid (BCA) assay, which is used to determine the total level of protein in a solution. Bicinchoninic acid is used to analyze to monitor cuprous ion presence due to its purple coloration via a biuret reaction.