Thursday, March 8, 2012

Agilent Offgel vs SCX

Overview:
~800 ug of peptides were divided in half
1 half went to SCX on a 10mM sodium phosphate (pH 2.8) gradient with an increase to 0.6M KCl.  20 fractions were collected
1 half was separated by peptide offgel using a high resolution strip pH 3-10
The peptides were desalted by ZipTips and loaded on our Velos using a standard top 10 method.
The MS/MS spectra were analyzed using Proteome Discoverer 1.3 with both Sequest and Mascot using Peptide Validator.
The results are shown above.  Pretty one-sided.  We'll see what the repeats look like

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