Does anyone know how a protein band can be extracted from SDS-PAGE gel without using any commercial kit?
The gel is stained by coomassie brilliant blue.
A razor blade. Just cut out your band by hand, put it in an eppendorf and dissolve the gel according to your needs. If you mean how to dissolve the gel, look at the manufacturing instructions, but acids, bases, peroxide, and other solvents commonly work.
Dear Bushra,
To extract the protein band from SDS PAGE, you have to do trypsin in-gel digestion of protein after cutting the protein bands. There is no protocol of dissolving the gel.
Follow this link in which there is a protocol for
Stage1: Excision of Coomassie stained bands
Stage 2: Reduction /Alkylation of band pieces
Stage 3: Digestion of band pieces
Stage 4: Extraction of peptides
http://www.lamondlab.com/pdf/LLingeldigestion.pdf
I hope this will help you. Good luck.
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