I am doing my project work on chick embryo as model. I want to find out what is the protein content in liver, muscle and brain tissue homogenates. While doing Bradford's assay, I wasn't getting Standard curve with BSA. I am adding protein stock, dilutes as HBSS, NaOH (0.25 ml) and bradford's reagent (5 ml).
What should I do? Is overall volume affected?
I never use NaOH when I do the Bradford assay. The dye is dissolved in acidic solution, so adding NaOH is probably unacceptable, although it would, of course, be concentration-dependent. If you need to add NaOH in your sample processing, neutralize it with an equivalent molar amount of HCl before doing the protein assay.
Perhaps the instructions say "NaCl" and not "NaOH". That would actually make much better sense.
Check this link maybe helpful
https://www.ncbi.nlm.nih.gov/m/pubmed/8509604/
regards
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