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CD45RA Antibody Staining Protocol for Immunohistochemistry
Description: The CD45R subfamily comprises a restricted form of the leucocyte common antigen and is divided into three isoforms: CD45RA, CD45RB and CD45RO. The CD45RA molecule, a 220kd isoform of CD45, is found on B cells, monocytes and a small proportion of T cells.
Primary Antibody
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Name: CD45RA Antibody |
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Clone: X148, Mouse anti-Human |
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Supplier: Novocastra Labs |
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Catalog Number: NCL-B1 |
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Dilution: 1:25 using IHC-TekTM Antibody Diluent (Cat# IW-1000 or IW-1001) to reduce background and unspecific staining and serum blocking step is NOT needed. |
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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: Mayer's Hematoxylin |
| Staining Time: 30 seconds |
Results:
| Staining Pattern: Membranes and cytoplasm of normal B-cells |
| 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 or 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: