TPCK-trypsin

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cavaliersun2003
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TPCK-trypsin

Postby cavaliersun2003 » Sun Oct 26, 2014 7:00 pm

Hey, all,
I'm wondering if anyone here have tried TCK-trypsin before. MS-grade trypsin, for example the one from promega, is used routinely but I think it is very expensive, especially when I use it to digest BSA just for instrument performance check. Is there a good quality but cheaper TPCK-trypsin alternative that can be used instead?
thank you!

SunSun :)

Kelstrup
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Postby Kelstrup » Mon Oct 27, 2014 2:09 am


Infinity
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Postby Infinity » Mon Oct 27, 2014 4:38 am

Order this one (cat #T1426) from Sigma, ~ 1000X less expensive and no difference. Completely replaced Promega trypsin in our lab.

cavaliersun2003
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Postby cavaliersun2003 » Mon Oct 27, 2014 7:36 pm

thank you!
however, T1426 is bovine trypsin.
Bovine trypsin were believed to be less specific than porcine, just like these two papers pointed out:
http://pubs.acs.org/doi/abs/10.1021/pr4001465
http://www.ncbi.nlm.nih.gov/pubmed/24116745
I'm wondering what specific digestion conditions (or treatments) you have used to make sure the bovine trypsin worked just like procine trypsin?

Infinity
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Postby Infinity » Tue Oct 28, 2014 5:56 am

The same conditions as for trypsin from Promega, side by side comparison revealed only very minor differences (we've got 3-5% less peptide IDs for T1426).

tsbatth
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Postby tsbatth » Tue Oct 28, 2014 12:21 pm

What trypsin to ptotein ratio are you using for your BSA? If it's just one protein you can get away with 1:200 or even 1:500 .

cavaliersun2003
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Postby cavaliersun2003 » Tue Oct 28, 2014 8:30 pm

tsbatth wrote:What trypsin to ptotein ratio are you using for your BSA? If it's just one protein you can get away with 1:200 or even 1:500 .

I will certainly try that out. Do you use longer incubation time for such diluted BSA:trypsin ratio?


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