B Lymphocyte Antigen 36kD Antibody Staining Protocol for Immunohistochemistry

Description: The B lymphocyte antigen, BLA.36, with a molecular weight of 36kD, is a membrane associated glycoprotein expressed on early progenitor B cells and B cells. BLA.36 antigen has a similar molecular weight to CD20 and CDw75 (LN1) but has been shown to be distinct from these haematopoietic cell lineage antigens. BLA.36 antigen may have a growth-related role during normal development of B lymphocytes. T lymphocytes, histiocytes, granulocytes, macrophages, stromal cells in lymphoid tissue and epithelial cells are all consistently negative for the expression of BLA36 antigen.

Primary Antibody

Name: B Lymphocyte Antigen 36 kD Antibody

Clone: A27.42, Mouse Anti-Human

Supplier: Novocastra Labs

Catalog Number: NCL-BLA36

Dilution: 1:25 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: Antigen retrieval is not needed

Buffer/pH value: N/A

Heat/Cool Temperature: N/A

Heat/Cool Time: N/A

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: Membrane

Images: Search image

Additional Information:

Tissue Type: Tonsil

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

Positive Control: Tonsil

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:

There is no need to perform antigen retrieval on acetone fixed frozen sections

References: