Hello
I am having a lot of difficulty with the Brownlee Spheri-5 PTH, 5 u, 220x2.1 mm
columns that we purchase from ABI for the protein sequencer.
The front half of the chromatogram is OK (separation and such) but the DTT is
high. The back half, Pro and further, is not good. The peaks are too small.
Proline is very large and tech support thinks that DPU is coming out too early
and coeluting with Pro. The PMTC peaks are also way small.
I am running Buffers A3 and B2. All reagents are fresh. New standards were
prepared
I tried putting back on my old column and these problems went away.
Unfortunately the column is too old to use now.
I have tried two new columns from ABI with the same result.
Any clues?
Thanks
Judy Phelps
phelp001@mc.duke.edu
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