RE: rat on mouse immunos

From:Cynthia Favara <>

	Depending on what tissue you are staining you may need only to use a
no primary control and a normal control. I do mostly CNS and the staining is
confined primarily to the blood vessels and is easily distinguished form
positive staining due to the primary antibody. I have used Innogenex mouse
on mouse kit which if I remember correctly biotinylates the primary before
application and eliminated need for the anti mouse secondary. This works
very well in my hands and is simple.
Cynthia Favara
Rocky Mountain Laboratories
903 S. 4th Street
Hamilton, MT 59840
FAX 406-363-9286

-----Original Message-----
From: Rita Angel []
Sent: Wednesday, March 28, 2001 1:49 PM
Subject: rat on mouse immunos

Hi histonetters,

I have an immuno question for the animal research people out there.  I have
a mac-3 antibody which is rat anti-mouse.  I will be staining mouse tissue.
  What kind of blocks do you use to block endogenous staining?  Our normal
block to use is 0.5ml of 2% mouse serum in 25mls of secondary.  I rotate
this for 1 hr. to allow binding and then use at the normal time (10 min.)
Does anyone else have any other procedure that they use?  This did not work
the last time I used it.

Thank you for your input,

Rita Angel

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