I have unknown peptide in known protein solution and I would like to isolate them for sequence analysis through MALDI MS/MS. I did in Diethyle ethar precipitation and found some problem during MALDI spot. It was forming a layer in MALDI plate (After drying of spot ). I have very few cystein in my protein. Therefore, DTT and IAD was not treated in solution. All the peptide solution was lyophilized before MALDI experiment with addition of 0.1% Formic acid. Please give suggestion to remove the barrier.
Hello Pritiprasanna Maity
It is not necessary to separate the protein and peptide. Unlike electrospray mass spectra, the MALDI-TOFMS is not complicated by multiply charged ions. That is a great advantage to 'look' at peptides in presence of proteins.
The responses of peptide and protein need not be linear in the spectrum. That is to say, one should not compare the peptide peak area to protein peak area to estimate the relative abundance.For the same sample/matrix, source parameters can be optimized for either the peptide or the protein.
We recently presented a poster at the MassSpec-2018 conference on similar topic.
Conference Paper Mass Spec Approaches to Detect Clipped Forms of Target-relat...
It is certainly true that the approach may not work in all cases, but give it a try.
Yes, It is possible provided you follow the recommended approach see article by Yekaguri etal 2005
Dr. Kaale, Sir., Thanks for your response. Can you please mention full title of the paper? So, I can find out the paper from google. Thanks again.
It is a poster presented at the MassSpec-2018 (CASSS) conference. Abstract can be seen at the ResearchGate link. Send me a request for private sharing of the poster.
Conference Paper Mass Spec Approaches to Detect Clipped Forms of Target-relat...
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