[Histonet] Cell Marque Mammoglobin (31A5) New Protocol?

[Truscott, Tom]

Hi Abby, Since the concentration is about a third of the original, I would try programming the XT to run the primary for 2 hrs instead of 32 min. You may also need to up the secondary time if that isn't sufficient. Tom T

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Abby L. Maples
Sent: Wednesday, January 22, 2014 6:11 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Cell Marque Mammoglobin (31A5) New Protocol?

Fellow histotechs,

We've been running mammoglobin testing on our Ventana Benchmark XT.
However, Cell Marque (our supplier of mammoglobin antibody) has changed the concentration. Our old concentration was 0.12 mg/mL, the new concentration is weaker at 0.04 mg/mL. The antibody is a rabbit monoclonal. We tried running the old protocol with the new concentration, but the results were not adequate (see below). We tried running the protocol given to us by Cell Marque, but theirs is specific towards the Benchmark UltraView and also returned inadequate results.

We process our tissue from formalin to paraffin. Our lab uses the iVew DAB detection kit.

Our old protocol:
Deparaffinization
Conditioner #1 (short 8 minute conditioning) Mild CC1 (mild 30 minutes conditioning) Standard CC1 (standard 60 minute conditioning) No enzyme 37C antibody incubation temperature No titration
Antibody: Mammoglobin 31A5 at 0.12 mg/mL concentration for 32 minutes A/B Block Hematoxylin counterstain 4 minutes Blueing post counterstain 4 minutes

Does anyone else use this product from Ventana/Cell Marque? Does anyone have a working protocol for a weaker antibody titration?

Any help would be greatly appreciated!

*Abby L. H. Maples, BS, MPH, HTL (ASCP)CM* Histotechnologist, Surgical Pathology Mercy Medical Center
701 10th Street SE
Cedar Rapids, IA 52403
Lab Phone: (319) 398-6827
Cell: (319) 432-1530
Fax: (319) 221-8767
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet