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| Protocols and Methods Forum Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique. |
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06-29-2008, 12:49 PM
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 TAP-TAG help Hey, i'm currently trying to do tap-tag purification on 2 proteins and seem to be somewhat struggling a little. Everything is fine until TEV cleavage. Upto now i've been incubating 10 units of TEV in 30ul TEV buffer for 5-10ul packed beads / mg extracted protein. overnight at 4 degrees... classic, yet it isn't as efficient as it should be. From the aliquotes it looks like cleavage is occuring, although at a low rate, but nothing is recovered in the supernatant... If anyone has had to deal with TEV cleavage, or has any technical advice, give me your best shot please  Uglyjelly        |
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07-01-2008, 07:51 PM
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 Re: TAP-TAG help forget it.. i found a solution. Just tried cleavage at room temp and it worked (plus my proteins didn't degrade)... Now that just means i'm not recovering proteins because they stick to the beads in a non specific manner... blablabla |
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| enzymatic reaction , purification , recombinant protein , taptag , tev proteasee |
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