I apologize for the basic question, but we are trying to do the BCA assay for the first and we can't seem to find what linear working range means. If you could also give any advice or describe how you perform BCA with microplates to have a reference (aside from the company's protocol) it would be much appreciated.
Dear Cristina Sanchez Cereceda
BCA assay is a calibration assay to determine the protein concentration. I have attached a manufacturer protocol.
In e-tube, ~6uL BCA+ 996 uL working dye (https://www.bio-rad.com/ko-kr/product/bio-rad-protein-assay?ID=d4d4169a-12e8-4819-8b3e-ccab019c6e13) 1+4ml 3DW. And pour 200uL from each standard and sample. Make a calibration curve from a range of concentrations of BCA standard.
https://assets.thermofisher.com/TFS-Assets/LSG/manuals/MAN0011430_Pierce_BCA_Protein_Asy_UG.pdf
Hello Cristina Sanchez Cereceda
BCA assay is used to determine the protein concentration. The protein concentration is determined with the help of a standard graph. It has different known concentrations of a reference protein, such as BSA against the absorbance values. The manufacturer mentions a linear working range, which means your reference concentrations within that range give a linear plot. It gives a more accurate value for your unknown sample concentration.
Microplate assay is also the same only difference is it's dealing with small volumes. Refer to the manufacturer's recommended range, but you can always optimise the methodology (incubation time, the volume of the reagent, buffer system used) according to your need. Hope this clears your doubt.
Best wishes!
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