Lawrence P. Casson
lpcasson at phoenix.Princeton.EDU
Thu Jul 8 22:43:12 EST 1993
In article <C9vH81.37G at biobase.aau.dk> ptk at biobase.aau.dk (Peter Kristensen) writes:
>I have been trying to get hold of some phage M-13 antibodies, but so far
>without much luck. I have contacted DAKO (Denmark), and they informed me that
>they once produced sheep anti-M13, but this product is no longer for sale.
Try: 5 Prime -> 3 Prime, Inc.
Boulder, CO 80303 USA
800-533-5703 (U.S. & Canada)
They sell cat# 5307-916192 Unconjugated Sheep Antibody (Polyclonal)
cat# 5307-187156 same as above, biotinylated
cat# 5307-648923 Biotinylated Donkey Anti-Sheep IgG
In my hands, both antibodies cross react to some extent with non-M13
material in bacterial phage supernatants. I have been using various
capture reagents to bind phage-antibodies, and detecting bound phage
with these reagents. I seem to get the least background signal when
I block the microtiter plates with BSA instead of milk. TWEEN increases
the background considerably. It also helps alot if I dilute my phage
supernatants 1/10 in PBS+1%BSA. I hope this helps.
I am looking for a monoclonal anti-M13 antibody since the reagents above
cross react with Donkey and Goat reagents that I could otherwise use for
my assays. My initial quick and dirty attempt at making a mouse
monoclonal was a failure due to insufficient purification of the phage.
I might try again with a better purification protocol. I am told that
CsCl purification removes many but not all contaminants. Unfortunately,
pillus proteins are among the ones that remain behind.
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