Blotting onto PVDF

Yong Gao ygao at NOVOPHARMBIOTECHINC.COM
Wed Aug 26 16:40:00 EST 1998


In replying to Karen Spink message "Blotting onto PVDF"

We routinely use the CAPS buffer that you mentioned to transfer proteins to
PVDF membrane for sequencing. It works very well. I would suggest you to
ckeck your protocol carefully. Have you soaked the PVDF membrane in 100%
Methanol for a few minutes prior to soaking in the transfter buffer? (if not
you won't get a transfer).

Yong




From: pdxkgs at pdn1.gene.nottingham.ac.uk (Karen spink)
Subject: Blotting onto PVDF
Date: Mon, 03 Aug 1998 09:58:46 +0100


Has anyone had any problems Western blotting onto PVDF in CAPS buffer? I am
trying to get a sequence on my protein and it appears that the efficiency
of transfer is letting me down. Could it be the buffer, (10mM CAPS, 10%
methanol, pH 11) ? I have had no problems blotting onto nitrocellulose
using tris-glycine buffers.

Thanks for your time

Karen Spink




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