CD54 (ICAM-1) Antibody Staining Protocol for Immunohistochemistry

 

Description: The intercellular adhesion molecule-1 (ICAM-1), also referred to as CD54, is an integral membrane protein of the immunoglobulin superfamily and plays a role in the cell adhesion. CD54 is found on resting (weak) and activated (moderate) lymphocytes and monocytes.

 

Primary Antibody

Name: CD54 (ICAM-1) Antibody

Clone: ICAM-1, Mouse anti-Rat

Supplier: BD Pharmingen

Catalog Number: 550302

Dilution: 1:50 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: Incubator
Buffer/pH value: IHC-TekTM Proteinase K Solution (Cat# IW-1101)
Heat/Cool Temperature: 37 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: Membrane/cytoplasmic of endothelial cells
Images: Search image

Additional Information:
Tissue Type: Tonsil, liver, kidney
Fixation: Formalin fixed paraffin sections, or acetone fixed frozen sections
Positive Control: Tonsil, liver, kidney
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

Notes:

Not need to perform antigen retrieval on acetone fixed frozen sections

 

References: