Re: CD31 protocol

From:Phyllis Davie

Sven,
    Here is the protocol we use for CD31.


CD31;  clone 1A10
Mfgr/Vendor:  Novocastra# NCL-CD31-1A10

Control:  tonsil 

Pretreatment:  Steam in 10mM citrate buffer, pH 6.0(+/-0.3) for 20 minutes
in vegetable steamer, followed by a 20' cooldown.   Wash in buffer.

Dilution:  Apply at 1:200 and incubate for 40 minutes @RT.    Wash in
buffer. 

Secondary:  biotinylated anti-MOUSE IgG, Vector # BA-2000, 1:300 in PBS
(incubate 20¹ @ RT) Wash in buffer.

Detection:  Elite  Avidin-Biotin-Complex‹Horseradish Peroxidase (ABCHRP),
Vector # PK-6100, made per kit instructions.  Incubate 20 minutes @RT.
Wash in buffer. 

Chromogen:  DAB


Best of luck!


Phyllis Davie
PhenoPath Laboratories--Seattle, WA
pdavie@phenopath.com



on 4/19/02 1:21 AM, Sven Terclavers at Sven.Terclavers@med.kuleuven.ac.be
wrote:

> Hi listers,
> 
> Does anyone of you know of a protocol to stain CD31 without the use of
> trypsine?!
> Greetz,
> 
> Sven
> 
> 





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