Bcl-6 Antibody Staining Protocol for Immunohistochemistry

Description: The bcl-6 gene, originally cloned from a tumor with 3q27 translocation, is commonly expressed in large cell lymphomas. In humans, bcl-6 encodes for a Kruppel-type zinc finger protein and is believed to be important in germinal center formation. Bcl-6 protein is expressed mainly by follicle center cells and in a few interfollicular T lymphocytes. Bcl-6 has also been detected in Burkitt’s lymphoma and in nodular, lymphocyte predominant Hodgkin’s disease. Bcl-6 is not expressed in hairy cell leukemia, mantle cell and marginal-zone derived lymphomas. Reactivity shows strong nuclear staining.

Primary Antibody

Name: Bcl-6 Antibody

Clone: PF16, Mouse Anti-Human

Supplier: Biocare Medical

Catalog Number: CM223B

Dilution: 1:50 using IHC-Tek^TM Antibody Diluent (Cat# IW-1000 or IW-1001) to reduce background and unspecific staining and serum blocking step is NOT needed.

Incubation Time/Temp: 60min/room temperature

Antigen Retrieval

Device: IHC-Tek^TM Epitope Retrieval Steamer Set (Cat# IW-1102)

Buffer/pH value: IHC-Tek^TM Epitope Retrieval Solution (Cat# IW-1100)

Heat/Cool Temperature: 95-100 ºC/room temperature

Heat/Cool Time: 20 minutes/20 minutes

Detection Methods

Standard Method: ABC Method or LSAB Method

Enhanced Method: Polymeric Methods

Chromogen Substrate

Reagent: DAB

Incubation Time/Temperature: 1-3 minutes/room temperature

Counterstain

Reagent: Mayer's Hematoxylin

Staining Time: 30 seconds

Results:

Staining Pattern: Nulcear

Images: Search image

Additional Information:

Tissue Type: Tonsil, follicular lymphomas

Fixation: Formalin-fixed paraffin sections

Positive Control: Tonsil, follicular lymphomas

Negative Control: Omit primary antibody, isotype control, absorption control

Blocking: 2-5% normal serum to reduce unspecific background staining; 0.5-3% H2O2 to block endogenous peroxidase activity; avidin/biotin to block endogenous biotin activity if necessary

References: