Re: How can I digest my target protein in-gel using Endoproteinase Glu-C enzyme

Dear Anas,

Thanks for your question. I ran a quick Google search for "protease V8 in-gel digest" and it threw up plenty of hits. One of them was:

Gallagher, MJ and Cohen, JB (1999).
Identification of amino acids of the torpedo nicotinic acetylcholine receptor contributing to the binding site for the noncompetitive antagonist [(3)H]tetracaine.
Mol Pharmacol. 56(2):300-307.
[PubMed] | [Full text]

The Experimental section states: "The band containing the nAChR alpha subunit was excised from the gel and placed on top of the stacking portion of a 15% mapping gel for in gel digestion with S. aureus V8 protease (Pedersen et al., 1986). This second reference (in J Biol. Chem.) is not freely-available, but should be accessible from your institution. It describes the V8 digestion in some detail.

If that doesn't help you, try the Google search or a Google Scholar search, as I did. There are very many protocols and procedures available on the Web these days, if you go out and look for them. Another excellent, growing resource to which you can even contribute is the OpenWetWare wiki.