I need to estimate protein from wheat grain/flour sample using the Bradford method. What is the standard procedure of extraction and estimation of protein using the Bradford method?
Dear Subhradip Bhattacharjee ,
Before starting I recommend you to read the following excellent overview related to protein determination:
Mæhre, H. K., Dalheim, L., Edvinsen, G. K., Elvevoll, E. O., & Jensen, I. J. (2018). Protein determination—method matters. Foods, 7(1), 5.
https://www.mdpi.com/2304-8158/7/1/5
It nicely describes the pros and cons of every method (including Bradford).
The consensus seems to be (especially in the field of cereals) that the Kjeldahl method is generally used as the standard method which can be compared with for example the Bradford method. Numerous papers describe the Bradford method in detail, see for example:
Kruger, N. J. (2009). The Bradford method for protein quantitation. In The protein protocols handbook (pp. 17-24). Humana Press, Totowa, NJ.
Article The Bradford Method for Protein Quantitation
Hope this helps you further.
Best regards.
PS. Do realize that in wheat different type/groups of proteins exists albumins, globulins, LMW and HMW glutenin’s that require different extraction procedures, see for a brief description of the various types of protein (fractions) for example:
Article Competitive Adsorption Behaviour of Wheat Flour Components a...
And for a very detailed description of the methods of extraction:
Tatham, A. S., Gilbert, S. M., Fido, R. J., & Shewry, P. R. (2000). Extraction, separation, and purification of wheat gluten proteins and related proteins of barley, rye, and oats. In Celiac Disease (pp. 55-73). Humana Press. (see enclosed file).
Dear Subhradip
Maybe, this article can help you:
Article Protein Determination-Method Matters
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