[Histonet] murine CD4, CD8 and CD68 for FFPE tissue

[Katy Milne]

Hi all,

I tried to respond yesterday but forgot to cut off a bunch making the email too big but I can add to the comments below...

First up, Gayle, thanks for the heads up that the CD8 is finally out, I'd heard it was happening and had been checking back but hadn't seen anything yet.  Will get that ordered ASAP.

Joseph I have used the CD4 and it looks pretty good so far, I'm not sure how to attach images on here but I can send you a stain although I've mostly been using it for multicolour stuff.  I use all Biocare reagents on an Intellipath FLX for my staining so we could see if we could translate a protocol to whatever platform you're using.

Joost, you can multiplex some of these Abs to get all your markers on one slide if you'd like.  I have pooled the CD4 mentioned and CD3 SP7 clone in the first round (which generally highlights CD8s as single positives vs the double positive CD4/CD3s) and then put F4/80 in the second round as a mac marker in place of CD68.  Didn't get back to fully fleshing it out yet but I also pooled PAX5 for B cells in there and got some nice images of tertiary lymphoid structures in mouse tissue.    We're doing some more staining with it today for a different project.

Anyway, CD4 Ab looks good so far, will order CD8 in for testing soon!

Katy

Message: 8
Date: Wed, 13 May 2015 18:12:49 +0000
From: Joseph Hardin <hardin at oncology.wisc.edu>
To: Histonet <histonet at lists.utsouthwestern.edu>,
	"gayle.callis at bresnan.net" <gayle.callis at bresnan.net>
Subject: Re: [Histonet] RE.....  murine CD4, CD8 and CD68 for FFPE
	tissue
Message-ID: <1431540771819.68363 at oncology.wisc.edu>
Content-Type: text/plain; CHARSET=US-ASCII

Has anyone tried these Abs? If so, please send me your beautiful photos and protocol.

Joseph Hardin
Senior Research Specialist
UWCCC Experimental Pathology
WIMR I Rm. 4012
1111 Highland Av.
Madison, WI 53705
(608)262-1836


________________________________________
From: Gayle Callis <gayle.callis at bresnan.net>
Sent: Wednesday, May 13, 2015 12:34 PM
To: Histonet
Subject: [Histonet] RE.....  murine CD4, CD8 and CD68 for FFPE tissue

You wrote:



Is anyone of you familiar with the possibility of applying CD4, CD8 and CD68 antibodies on formalin fixed paraffin embedded liver of mice?



Thanks in advance.

Joost Bruijntjes



**************************************

Thankfully and at long last after years of frustration by many ................



eBioscience has both the CD8a and CD4 which is now available for FFPE murine tissues.  Read the data sheets carefully as the CD4 indicated this antibody was to be used on FFPE and not frozen sections.

            Anti-Mouse CD8a Purified Catalog Number: 14-0808 Also known as:
CD8 alpha, Ly-2, Ly-35, Lyt-2

            Anti-Mouse CD4 Purified Catalog Number: 14-9766 Also known as:
L3T4, Ly-4



Serotec has a rat anti mouse CD68 where the data sheet says can be used on FFPE murine tissue.  This clone must be available from other companies as well.



Gayle M. Callis

HTL/HT/MT(ASCP)



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