The Bicinchoninic Acid (BCA) Protein Assay is a highly sensitive colorimetric assay that is compatible with detergent solubilized protein solutions, however is dramatically affected by the presence of reducing agents, including DTT, ß-mercaptoethanol and TCEP.
The Bicinchoninic Acid (BCA) Protein Assay primarily relies on two reactions. Firstly, the peptide bonds in the protein sample reduce Cu2+ ions, in a temperature dependent reaction, from the copper solution to Cu+. The amount of Cu2+ reduced is proportional to the amount of protein present in the solution. Next, two molecules of bicinchoninic acid (BCA) chelate with each Cu+ ion, forming a purple-colored product that strongly absorbs light at a wavelength of 562 nm that is linear for increasing protein concentrations between the range of 0.02-2mg/ml.
The Bicinchoninic Acid (BCA) Reducing Agent Compatible Protein Assay is supplied with the Reducing Agent Compatibility Agent (RACA) that modifies reducing agents to limit their effect on the reduction of the assay’s copper ions, preventing inhibition of the assay. The use of RACA allows for samples containing up to 5mM DTT, 10mM TCEP or 35mM ß-mercaptoethanol.
The Bicinchoninic Acid (BCA) Reducing Agent Compatible Protein Assay is suitable for quantifying protein solutions in 1ml assays or in micro-wells. This kit is suitable for is for 250 x 1ml assays.
- Reducing agent compatible assay
- Sensitive colorimetric assay
- Linear range: 0.02-2mg/ml
- Compatible with most ionic and non-ionic detergents
- For measuring protein concentration in samples with reducing agents