Western Blotting of small proteins

Vuk Stambolic stambo at ocicl.oci.utoronto.ca
Thu Sep 26 11:29:15 EST 1996


A friend of mine that works in Italy but has no access to this newsgroup
has a following question:

 I want to try and detect a truncated protein which is found
in certain cancer cells. I have good antibodies to the n-terminal and can
see the native protein...no problem. I have taken the cancer cells and run
the proteins on a gel (17% PAGE (also a 12.5% with a 20% step)), but see
nothing after western blotting (transfered to PVDF). I realise that PVDF is
good for proteins >10K. My truncated protein is probably in the 5-6K range.
Has anyone had success in detecting such small proteins by western blotting
or I am I loosing the protein. If so, is there a better membrane with a
smaller pore size (<0.45 micron) that someone has used succesfully for
western analysis of small proteins.

Please reply to him directly at:
plyte at dada.it (Simon E Plyte)

Thanks,
Vuk



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