RE: Mouse in mouse

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From:Cynthia Favara <cfavara@niaid.nih.gov>
To:"'Histonet'" <HistoNet@Pathology.swmed.edu>, "'Blanca Arango'" <blanca_arango@yahoo.com>
Reply-To:
Date:Fri, 16 Apr 1999 12:25:41 -0400
Content-Type:text/plain

Blanca,
	Do you have to use both antigen retrievals?
	Have you tried diluting your secondary further or decreasing the
temp or time?
	For mouse on mouse I dilute my secondary in TBS wtih 5%skim milk
seem to help with the background.

Good Luck

Cynthia Favara
Rocky Mountain Laboratories
903 S 4th Street
Hamilton, MT 59840
ph: 406-363-9317
FAX: 406-363-9286
e-mail: cfavara@nih.gov


> ----------
> From: 	Blanca Arango[SMTP:blanca_arango@yahoo.com]
> Sent: 	Friday, April 16, 1999 8:51 AM
> To: 	'Histonet'
> Subject: 	Mouse in mouse
> 
> Hi Histonetters,
> I am looking for the expression of p27-protein in mouse retina using an
> anti-p27 MOUSE-antibody. To develop the reaction I use the DAB system,
> but I find false positives in the control slides (without first
> antibody). I work in formalin-fixed, plastic embedded tissue sections.
> My protocol includes trypsin digestion and microwave antigen retrieval.
> I have already
> tried endogenous peroxide activity blocking and endogenous biotin
> blocking. 
> Does anyone have a solution?
> Does anyone have any experience in blocking of endogenous Ig?
> 
> Thanks for your input.
> 
> Blanca Arango Gonz#225#lez 
> University Eye Hospital Tuebingen
> Experimentelle Ophthalmology
> Roentgenweg 11 
> D-72076 Tuebingen 
> Germany   
> Tel.: +49 - 07071 - 29 87425 
> Fax: +49 - 07071 - 29 5777
> 
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> 



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