ECL background, addendum

Helmut Dotzlaw dotzlaw at ccu.umanitoba.ca
Thu Jul 29 18:47:02 EST 1993


Good day, all,

Tried something different today and was impressed by how well it worked, so
thought to share.  A colleague suggested a variation on Amersham's ECL
detection to minimize the background on long exposures, and also to use
less reagent per blot. 

Following the washes after the secondary Ab, take the blot out of TBST and
pat it "damp" (no puddles) with some tissue, then place it protein-side
down on a piece of plastic wrap.  Layer the reagent mix (2 ml for a 4x8 cm
minigel) on top of the blot and wait 1 minute.  Pat dry again, wrap, and
expose to film.

The result - a 1 hour exposure with absolute zero background, a "first" for
me!

BTW, I used Nitroplus 2000 nitrocellulose membrane, not a "plug", just what
I have.

Regards,

Helmut



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