Is urea compatible with TMT/iTRAQ reagents?

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gabe
Angiotensin Member
Angiotensin Member
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Joined: Sat Nov 19, 2011 5:56 pm

Is urea compatible with TMT/iTRAQ reagents?

Postby gabe » Mon Dec 17, 2012 11:27 am

I know that TMT tags are not compatible with buffers containing primary amines (like Tris or ammonium bicarb) but I was wondering about urea: chemically speaking, I think the urea amides should be pretty inert and should not interfere with the NHS chemistry. Does anyone know if this is, in fact, correct? Is it necessary to remove all traces of urea from a solution digest before proceeding with the TMT reaction?

Thanks

alex
Carbon Member
Carbon Member
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Joined: Tue Dec 04, 2012 9:19 am

Postby alex » Mon Dec 17, 2012 1:52 pm

I have labeled in molar amounts of urea and haven't had problems. I would be wary of any heating of the sample, since urea degradation products are likely not compatible with labeling.

Biomarker
Albumin Member
Posts: 88
Joined: Thu Aug 09, 2012 3:28 am

Postby Biomarker » Tue Dec 18, 2012 12:09 am

Hi,

We never know how things will interfere, so its good to remove it.
I have done urea (used almost 4M) based trypsin digestion and desalted the peptides using Sep-Pak. Followed by labeling of desalted samples with 8 plex iTRAQ reagents. And i have not faced any difficulty in labeling. The labeling efficiency was almost 95%.

Biomarker.


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