Whenever I can I like to dilute standards with the same buffer used for my sample to avoid matrix effect. Also, if you work frequently with one protein it would be useful to have a standard composed of the protein you want to measure rather than BSA; in this case accurate protein concentration of the standard should be determined by amino acid analysis.
Dear Heba,
Check this protocol attached below
During my PhD work I used PBS to dilute
I hope this will help
http://ergo.berkeley.edu/be115/BCA.pdf
The BSA standards can be diluted with water. For my BCA assays, I dilute the standards to final concentrations: (0, 250, 500, 1000, 1500 and 2000μg/m
L) to generate a standard curve and determine an absorbance- concentration relationship. Some researchers plot a linear regression which unfortunately is inaccurate. I prefer using the curvilinear regression to determine unknown protein concentrations of my samples instead.
This technical tip from Thermoscientific should give you better insight:
https://tools.thermofisher.com/content/sfs/brochures/TR0057-Read-std-curves.pdf
Whenever I can I like to dilute standards with the same buffer used for my sample to avoid matrix effect. Also, if you work frequently with one protein it would be useful to have a standard composed of the protein you want to measure rather than BSA; in this case accurate protein concentration of the standard should be determined by amino acid analysis.
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