Re: Immunostaining Mouse Liver

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From:Jamie Erickson <JErickson@genetics.com>
To:roverj@earthlink.net, histonet@pathology.swmed.edu
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PHARMINGEN is your company, I've stained mouse liver, spleen, nodes you name it with Parmingens, CD45R/B220 and it states on the sheet paraffin embeded sections that usally means formalin fixed, you must have missed it. I've also used sertocs Rat anti- human CD3 on mouse paraffin tissues, I use  Proteinase K (25 mg/mL) for 20 ( for B220) minutes at 37 C as a enzyme digestion. Dedending on your tissues  fixation time you will need to adjust the enzyme time you'll have to work that out all my samples are fixed for 24 hours no more. Hope this helps.



Jamie Erickson
Associate Scientist
Genetics Institute
1 Burtt Rd.
Andover, MA   01810
work : (978) 247-1348
FAX  : (978) 247-1333

>>> Trevor Jorgensen <roverj@earthlink.net> 02/24 9:26 AM >>>
We are interested in staining formalin fixed, paraffin embedded mouse
liver for B-cells and T-cells.  While monoclonals are available for
frozen sections, (pharmingen among others) we have not located any which
are recommended for use with fixed material.
Has anyone had any experience (preferably with CD3 and CD19 or CD20 ) in
this situation?  How about with DAKO's ARK and using mouse against
mouse?






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