Re: CD31 protocol

From:Phyllis Davie

    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

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

on 4/19/02 1:21 AM, Sven Terclavers at

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

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