CD54 (ICAM-1) Antibody Staining Protocol for Immunohistochemistry


Description: Cell adhesion molecules are a family of closely related cell surface glycoproteins involved in cell-cell interactions during growth and are thought to play an important role in embryogenesis and development. Neuronal cell adhesion molecule (NCAM) expression is observed in a variety of human tumors including neuroblastomas, rhabdomyosarcomas, Wilm’s tumors, Ewing’s sarcomas and some primitive myeloid malignancies. The intercellular adhesion molecule-1 (ICAM-1), also referred to as CD54, is an integral membrane protein of the immunoglobulin superfamily and recognizes the B2A1 and B2AM integrins. PECAM-1 (platelet/endothelial cell adhesion molecule-1), also referred to as CD31, is a glycoprotein expressed on the cell surfaces of monocytes, neutrophils, platelets and a subpopulation of T cells. VCAM-1 (vascular cell adhesion molecule-1) was first identified as an adhesion molecule induced on human endothelial cells by inflammatory cytokines such as IL-1, tumor necrosis factor (TNF) and lipopolysaccharide (LPS). The KALIG gene encodes a nerve cell adhesion molecule (NCAM) -like protein and is deleted in 66% of patients with Kallmann's syndrome, anosmia with secondary hypogonadism.


Primary Antibody

Name: ICAM-1 Antibody

Clone: Goat anti-ICAM-1

Supplier: Santa Cruz Biotechnology

Catalog Number: sc-1511

Dilution: 1:200 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: IHC-TekTM Epitope Retrieval Steamer Set (Cat# IW-1102)
Buffer/pH value: IHC-TekTM Epitope Retrieval Solution (Cat# IW-1100)
Heat/Cool Temperature: 95-100 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

Reagent: Mayer's Hematoxylin
Staining Time: 30 seconds

Staining Pattern: Membrane/cytoplasmic of endothelial cells
Images: Search image

Additional Information:
Tissue Type: Tonsil, liver, kidney
Fixation: Formalin fixed paraffin 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