RE: mouse monoclonal antibodes.Sorry!!!!

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From:Matthew Ogdie <>
To:"''" <>
Date:Wed, 09 Jun 1999 12:52:04 -0700
Content-Type:text/plain; charset="us-ascii"

You were referring to Andrea's question - my apologies.

Matthew Ogdie

From: 	Gayle Callis[]
Sent: 	Wednesday, June 09, 1999 9:09 AM
Subject: 	mouse monoclonal antibodes

Oops, misspell!  

These clones of mouse  cell surface markers are tough.

Other companies also sell these specific clones, but they probably still
work best on frozens, consequently nothing has changed per trying to use
another vendors monoclonal, the problem remains.. and is the nature of the
beast, that little critter, mouse!

The CD45R/B220 will work on NBF,paraffin without pretreatment or use
an enzyme digestion.  It also works on Carnoys fixed tissue, no pretreatment.

PharMingen does not recommend paraffin for any of the others, but some
people may have had success with enzyme pretreatment, and overnight cold
incubation of antibody.  PharMingen is also recommending a zinc fixative,
NOT ZINC FORMALIN, this works well with many cell markers, and you may 
want to try it, morphology is pretty good, cell surface marker staining
is nice. Formalin is just not user friendly for mouse markers.   

One gentleman dropped me a file attachment, but have not heard from him
again (couldn't open it on work computer, hoped he would send it to me
at home address!!!) with successful staining on cell surface markers, mouse.
Hope he is out there looking in, waiting with bated breath.  
I also have a list of enzyme pretreatments recommended and compiled by a H'netter asking 
similar questions. (I thank him for putting it all together!) 

Guess this NBF/paraffin dilemma has kept us using acetone fixed frozen
sections, saves us lots of time, and always works for same markers. 

Give that fixative a try, tissues are a tidge drier, but workable.  You also
MUST use alkaline phosphatase kits, since endog peroxidase cannot be quenched
after this zinc fixative.  I know of several people who use it with great

Gayle Callis

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