[Histonet] CD31 problewms

[Rene J Buesa]

If you have not changed the Ab provider nor any step on your protocol, check the Ig concentration in the lot you are using now compared with the lot you used to determine the Ab dilution. If the Ig is less concentrated now, you may need to increase the concentration (reduce the dilution).
René J.
 

From: "Jo-Ann Bader, Ms." <jo-ann.bader <@t> mcgill.ca>
To: "histonet <@t> lists.utsouthwestern.edu" <histonet <@t> lists.utsouthwestern.edu> 
Sent: Thursday, February 14, 2013 11:49 AM
Subject: [Histonet] CD31 problewms

We are having problems getting our CD31 to work well on our NBF fixed mouse tissue. There is staining but it is very pale.

The CD31 comes from Bio-care and we are are using the IntelliPATH stainer.

We have tried:

- Trypsine 37C 15 min / Trypsine RT 30 min
- Trypsine (1:2 and 1:3)
- With Decloaker / without decloaker
- DAB Sparkle / No DAB sparkle
- Different dilution of CD31 (1:50 / 1:25)
- Different time of CD31 incubation (2h / 3h)
- Different time of incubation of secondary and tertiary reagent (10min to 20min)

Thanks


Jo-Ann Bader
Histology Co-Ordinator
Goodman Cancer Centre
McGill University 1160 Pine Ave. W - Rm 312
Montreal, QC, Canada
H3G 1Y6
Tel: 514-398-8270
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