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

Name: CD45RA Antibody

Clone: X148, Mouse anti-Human

Supplier: Novocastra Labs

Catalog Number: NCL-B1

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.

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

Reagent: Mayer's Hematoxylin
Staining Time: 30 seconds

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