I am planning to use high pH reversed-phase peptide fractionation kit from pierce (Catalog no.84868) for fractionating labelled peptides. I have used off-gel fractionator (Agilent) to fractionate the samples in the past and the results have not been great.
Any reviews for the pierce kit based on personal experience that could be of use to me ?
Hi there, what kind of labels for your samples? iTRAQ or TMT or any other labelling kits? I'm currently working on membrane proteins and I label my sample with iTRAQ. Previously, I did OFFGEL too but due to some problems with our fractionator, we are struggling to decide which peptide fractionation are capatible with iTRAQ labeled peptides. We also planned to use the Pierce high pH reversed-phase peptide fractionation kit, but we are not confident whether it is compatible with iTRAQ labeled peptides as it recommends for TMT labelled peptides.
Hi there, I used recently the high pH reversed-phase peptide fractionation kit from Pierce on non-labelled peptides. In my system, with non-fractionated samples, I identified between 2200 to 3000 proteins with 1% FDR (with ProteinPilot software). Using the pH fractionation kit, I have identified around 5900 proteins (1% FDR). Quite nice! The kit is validated for TMT labelling (from Thermofisher as Pierce!). I want to test it with iTRAQ also! The Technical Sales Specialist said to me that there will work.
It is comparable with the with our offline LC system but we have to compensate by increasing the gradient time for the analyses.
the stated capacity for the kit is " 10–100 µg of peptide ".
anyone knows of other commercially available kit with larger capacity? Reason being i am using UPLC instead of nanoLC, hence requiring more samples
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