iTRAQ and RapiGest

Share protocols and ask for sample preparation advice.
Proton Member
Proton Member
Posts: 1
Joined: Tue Dec 11, 2012 4:08 pm

iTRAQ and RapiGest

Postby juli_f » Tue Dec 11, 2012 4:24 pm

I 'd like to know if I can use RapiGest ressuspended in 50mM of NH3HCO4 to lysis my tissue and after digestion with tripsin, can I use iTRAQ?

Is RapiGest (ressuspended in NH3HCO4) compatible with iTRAQ?

Thank you in advance,

E. Coli Lysate Member
E. Coli Lysate Member
Posts: 107
Joined: Wed Dec 21, 2011 8:22 pm

Postby Infinity » Wed Dec 12, 2012 8:57 am

Well, I'm not sure about RapiGest, but ammonium bicarbonate is not compatible with iTRAQ since it contains amino groups (in the form of ammonia). It is recommended to desalt your peptides and use iTRAQ-compatible buffer (for example truethylammonium bicarbonate).

Angiotensin Member
Angiotensin Member
Posts: 42
Joined: Thu Dec 27, 2012 12:26 pm

Postby Christopher » Sun Dec 30, 2012 5:48 am

I have not tried it with iTRAQ, but RapiGest is compatible with TMT labeling (at 0.1%) in my hands. As above, you cannot use ammonium bicarbonate as your buffer...we use HEPES, but others are ok as well.

Return to “Sample Prep”

Who is online

Users browsing this forum: No registered users and 1 guest