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

[Abby L. Maples]

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