iTRAQ and RapiGest

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juli_f
Proton Member
Proton Member
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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,
Juliana

Infinity
E. Coli Lysate Member
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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).

Christopher
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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.


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