help with block of PVDF

Jim Hutchins hutchins at fiona.umsmed.edu
Mon Nov 20 21:17:30 EST 1995


Stefan Bäckström (stefanba at bio.embnet.se) wrote:
: What do you use for blocking PVDF membranes ?

1. 1-2% Blotto(tm) in PBS or TBS (Carnation Non-fat Dry Milk, I'm sure any
   brand will do)...note milk contains phosphotyrosine residues so it's
   not suitable for all blots

2. 1% bovine serum albumin in PBS or TBS

3. 1% fish gelatin (Sigma) in PBS or TBS

4. 0.05% Tween-20, alone or in combination with any of the above

For low background, keep the secondary antibody concentration as low
as possible.  To reduce "extra" bands, reduce the protein concentration
in the gel...25 ug seems to be the 'magic' starting point.

Good luck
--
Jim Hutchins
Assoc Prof Anatomy, Asst Prof Neurology (Research), Univ Mississippi Med Ctr
http://fiona.umsmed.edu/~hutchins/    ***    E-mail:   hutchins at umsmed.edu
``It became necessary to destroy the town in order to save it.''--American
infantry officer firing on Ben Tre, Vietnam, 2/8/68



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