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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