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
