I’m using the Pierce® BCA Protein Assay Kit − Reducing Agent Compatible from Thermo Scientific, and I have a few questions.
Could you please refer to the protocol and help clarify the following:
https://assets.thermofisher.com/TFS-Assets/LSG/manuals/MAN0011538_Pierce_BCAProtein_Asy_ReducAgntComp_UG.pdf
Can I skip using the Compatibility Reagent and follow the same procedure as with other Pierce® BCA Protein Assay Kits (those without a Compatibility Reagent)? Would skipping the Compatibility Reagent affect the sample-to-working-reagent ratio? If I don't use the Compatibility Reagent, does that mean I only need to prepare one blank (the standard blank) instead of two (standard blank and sample blank) as required in this protocol?
Thank you
If you don't use Compatibility Reagent then Standard Blank = Sample Blank, You can skip Sample Blank and step 2.
DEar Wajd Alali
since the BCA is based on conversion of Cu(II) to Cu(I) is not compatible with the common reducing agents (GSH, DTT, B-me, TCEP) in mM concentration. Is possible that it can work in uM concentration but you have to test it in the blank.
Generally Bradord assaay or Absorbance measurment at 280nM is preferable as protein/antibnody quantification methods in presence of reducing agents.
best
Manuele
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