 Methods & Techniques For Histologists and Immunohistochemists |
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
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Name: B Lymphocyte Antigen 36 kD Antibody |
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Clone: A27.42, Mouse Anti-Human |
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Supplier: Novocastra Labs |
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Catalog Number: NCL-BLA36 |
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Dilution: 1:25 |
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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: Gill's Hematoxylin or 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: