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

 

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