Presently, I am using the Lowry method to estimate the protein concentration, however, I need to practice the Bradford method. As you all know that it entails Coomassie Brilliant Blue G-250 dye and cost is less expensive if I buy Coomassie blue dye (from sigma) and prepare the solution for Bradford assay rather than buying readymade solution. Since I don't have any previous experience in this experiment, I expect your suggestions about this, i.e. is ready-made solution superior than prepared one, are there any precautions to be considered before manual preparation, etc.
Thanking you in advance for your suggestions.
A precisely prepared Bradford solution is equivalent to a readymade solution purchased from BioRad or sigma.
Regarding precaution :
Double filter your solution using whatman filter paper No. 1.
Use HPLC grade ethanol and ortho-phosphoric acid and miliQ grade water
Store the solution in brown colored bottle and prepare around 400 ml that can be used up within 3 month.
Attaching a chapter on Bradford assay (by Prof. Kruger). I hope it to be useful in understanding the assay
Hai Sir,
I have one more doubt that, it has been generally instructed that protein standards should be prepared in the same buffer as the samples to be assayed. For instance, in Lowry method, protein standard is being prepared with NaOH solution. Hence, for Bradford assay, i have to prepare the protein standard with which solvent?.
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