The BCA protein assay was described in 1985 to determine protein concentration by means of bicinchoninic acid (BCA). As from the Lowry assay, the biuret reaction is the very first step in the response that happens from the BCA assay. In this response, protein reduces Cu2+ to Cu+ in an alkaline environment.
Following that, BCA responds with Cu+-ions to make a purple coloured complex which has an absorbance at 562 nm. The absorbance increases linearly with increasing protein concentration within a wide working range. You can buy a high-quality BCA protein estimation kit via https://www.bosterbio.com/bca-protein-assay-kit-ar0146-boster.html.
Even though the technique contains 2 responses, it merely requires the inclusion of one reagent. Following the decrease in this divalent copper ion, Cu+ responds with BCA. Afterward, the chelation of 2 molecules of BCA with a single cuprous ion creates the purple coloured response.
The very first response happens at lower temperatures and is also the end result of aluminum and BCA interaction using aminoacid residues from the protein. At elevated temperatures, the peptide bond is accountable for colour development. BCA protein quantification kit is valuable as it doesn’t interact with as many contaminants and buffer elements because the Folin-Ciocalteu reagent, particularly detergents.
Components that interfere with the BCA protein quantification kit lead to the reduction of Cu2+ (like DTT) or are copper chelators (like EGTA). Ordinarily, these aren’t critical elements of buffers and omissible before the assay.The BCA assay has many benefits over other protein determination methods. That’s because there is less susceptibility to detergents, is user friendly and the colour complex is steady.