Hallo Everyone,
We Have in our lab proteomic grade Trypsin Sigma Catalog Number T6567. AS per the product information sheet it is recommended that product stability is 1 year if stored at 4 degree Celsius.( https://www.sigmaaldrich.com/content/dam/sigma-aldrich/docs/Sigma/Bulletin/t6567bul.pdf ) And now it is almost after 3 years now I want to use the trypsin.I want to know whether the Trypsin still efficiently working?How to check the trypsin digestion efficiency without going for LC-MS analysis?or whether to go for buying New trypsin?Also I have Seen many papers where sequencing grade Trypsin with modified lysin (Promega) is used for better in-solution trypsin digestion..
1.So any one can help which is the best trypsin for in solution protein digestion proteomic grade Trypsin Sigma or sequencing grade Trypsin with modified lysin (Promega)( https://www.promega.in/products/mass-spectrometry/proteases-and-surfactants/sequencing-grade-modified-trypsin/?catNum=V5111 )
2.I want to know whether the Trypsin still efficiently working?How to check the trypsin digestion efficiency without going for LC-MS analysis?
Please reply...
Dear Sagar Swami,
I think your trypsin is probably working despite the time in the refrigarator.
Even though you can test is efficiency by digesting any protein such as bovin serum albumin or any other purified protein you have in your lab. you can see the result (the digested protein in form of bands) by electrophoresis.
Another more complicated approach is to perform a "active site titration". See the article:
Active site titration of bovine P-trypsin by W-(N,N-dimethylcarbamoyl)- a-aza-lysine p-nitrophenyl ester: kinetic and crystallographic analysis . FEBS Letters 358 (IY95) 53-56. There are other articles you can read about trypsin active site titration.
Regards,
Nelder
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