CD79a (B Cell) Antibody Staining Protocol for Immunohistochemistry

 

Description: Anti-CD79a is to identify B-cell linage (but not in myeloid or T cell lines) in formalin-fixed, paraffin embedded normal and neoplastic tissues using immunohistochemical (IHC) test methods. The CD79á polypeptide appears early in B-cell maturation at the pre-B-cell stage, and persists until the plasma cell stage, where it is found as an intracellular component. Anti-CD79á can be used for the immunohistochemical identification of cells of the B-cell lineage.

Primary Antibody

Name: CD79a Antibody

Clone: JCB117, Mouse anti-Human

Supplier: Dako

Catalog Number: M7050

Dilution: 1:100 using IHC-TekTM Antibody Diluent (Cat# IW-1000 or IW-1001) to reduce background and unspecific staining and serum blocking step is NOT needed.

Incubation Time/Temp: 60 min/room temperature

Antigen Retrieval

Device: IHC-TekTM Epitope Retrieval Steamer Set (Cat# IW-1102)

Buffer/pH value: IHC-TekTM 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: Cytoplasmic

Images: Search image

Additional Information:

Tissue Type: Tonsil, thymus, spleen

Fixation: Formalin fixed paraffin embedded sections, or acetone fixed frozen sections

Positive Control: Tonsil, thymus, spleen

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

Notes:

Not need to perform antigen retrieval for acetone fixed frozen sections

 

References: