I'd second what Tim says, and add that if background is still a problem
after using casein (non-fat dried milk) you could add either gelatin or PVA
(both 2%) to your blocking solution.
I would also like to mention to anyohne doing IF that, in Dallas, we
switched all our fluorescent secondaries to Alexa dyes (Molecular Probes),
as we get a much brighter signal,less fading on exposure to UV, and less
background. In fact we exposed a section with Alexa 594 (the equivalent to
Texas Red) continuously for 48 hours with very little fading of the
reaction (unpublished results).
Regional Histology Operations Manager
Bon Secours Health Partners Laboratories
5801 Bremo Road
______________________________ Reply Separator _________________________________
Subject: RE: Immunohistochemistry
Author: "Morken, Tim" at BSHSIBTW
Date: 4/12/02 9:15 AM
Solutions of powdered milk have been used (the protein casien
non-specific binding)and some companies sell their own
protein-free blocking agents.
From: Miller, Barbara [mailto:firstname.lastname@example.org]
Sent: Friday, April 12, 2002 8:48 AM
Cc: Miller, Barbara
Does anyknow of blocking agents other than serum or albumin
provide a low background for immunohistochemistry. We are
fluorescent dye (often Texas Red) and background is a problem so far.
Thank you for your help.
Department of Gastroenterology
Roudebush Medical Center
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