[Histonet] CD31 problewms

[Connolly, Brett M]

I think the CD31 Ab from Dianova is what works best for most people on FFPE mouse tissue. 

Brett

Brett M. Connolly, Ph.D.
Principal Scientist, Imaging Dept.
Merck & Co., Inc.
PO Box 4, WP-44K
West Point, PA 19486
brett_connolly <@t> merck.com
T- 215-652-2501
F- 215-993-6803





-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Rene J Buesa
Sent: Thursday, February 14, 2013 12:27 PM
To: Jo-Ann Bader, Ms.; histonet <@t> lists.utsouthwestern.edu
Subject: Re: [Histonet] CD31 problewms

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
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Notice:  This e-mail message, together with any attachments, contains
information of Merck & Co., Inc. (One Merck Drive, Whitehouse Station,
New Jersey, USA 08889), and/or its affiliates Direct contact information
for affiliates is available at 
http://www.merck.com/contact/contacts.html) that may be confidential,
proprietary copyrighted and/or legally privileged. It is intended solely
for the use of the individual or entity named on this message. If you are
not the intended recipient, and have received this message in error,
please notify us immediately by reply e-mail and then delete it from 
your system.