RE: [Histonet] blocking solution

From:"Favara, Cynthia \(NIH/NIAID\) [E]"

In my hands BSA often causes more problems than it solves. One needs to
be careful of the grade used, it is very 'sticky' and I have had more
non-specific background staining using BSA than with just about anything
else. We previously have studied mostly rodent brain and eye which I
have found to have very little background problems. Also if you are
using and goat primary you can get some cross-reactivity with BSA and
some anti-goat IgG's depending on purity, specificity etc. 

So to get to the heart of the matter I only block for endogenous
peroxidase unless I have a problem and then I will block with normal
serum of the host species of the secondary. I am becoming a minimalist!

c

Cynthia Favara
NIAID/NIH/RML/LPVD
903 South 4th Street
Hamilton, MT 59840
406-363-9317

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-----Original Message-----
From: Lexus Ray [mailto:spicin82@hotmail.com] 
Sent: Tuesday, May 16, 2006 12:03 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] blocking solution

Hi,
I'm a novice to immunolabeling.
What are the advantages /disadvantages of using BSA in a blocking
solution?
Supposedly I used only normal sera in my blocking solution, would this 
result in high background resulting from primary antibody?

L

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