I have to quantify a mixture of proteins. But I can not quantify its concentration by any protein detection methods as the values fluctuate. Can i quantify the protein concentration by sds page?
Hello Sreyasi
You can quantify protein bands in SDS-PAGE but by using densitometry with molecular weight marker of known concentration.
You can refer to the research article below :
Microbiologyopen. 2020 June, 9(6): e1027.
doi : 10.1002/mbo3.1027
Also please refer to one more research article below :
Integrated protocol for reliable and fast quantification and documentation of electrophoresis gels. Dec 2014
doi:10.1016/j.pep.2014.12.006
Regards.
Ideally, to determine total protein concentration are methods such as Bradford or else those based on fluorometry. Something less precise may be by absorbance.
Depending on how many proteins you have in your extract, various softwares (if you don't have a densitometry equipment) that allow estimating the concentration of the bands in an SDS-PAGE.
To do this you need to place a standard of concentrations of a protein of which you know the concentration (for example BSA) and then the band that interests you is compared. One of those software is called TotalLab, but there are others.
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