Integrin beta4 Antibody Staining Protocol for Immunohistochemistry


Description: Integrins are heterodimers comprising non-covalently associated, transmembrane and subunits. The sixteen and eight subunits heterodimerize to produce more than 20 different receptors. Most integrin receptors bind ligands that are components of the extracellular matrix, including fibronectin, collagen and vitronectin. Certain integrins can also bind to soluble ligands such as fibrinogen, or to counter-receptors on adjacent cells, such as the intracellular adhesion molecules (ICAMs), leading to aggregation of cells. Signals transduced by integrins play a role in many biological processes, including cell growth, differentiation, migration and apoptosis. Laminin receptor (Laminin-R) has a heterodimeric structure similar to that of receptors for other extracellular matrix proteins such as fibronectin and vitronectin and is a member of the integrin family of cell adhesion receptors. Incorporation of Laminin-R into lysosomal membranes makes it possible for lysosomes to attach to surfaces coated with laminin.


Primary Antibody

Name: Integrin beta4 (CD104) Antibody

Clone: ASC-3, Mouse anti-Human

Supplier: Chemicon

Catalog Number: MAB2058Z

Dilution: 1:100 for paraffin sections and 1:200 for frozen sections 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: 60 min/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

Reagent: Mayer's Hematoxylin
Staining Time: 30 seconds

Staining Pattern: Basement membrane
Images: Search image

Additional Information:
Species Reactivity: Human
Fixation: Formalin fixed paraffin sections, or acetone fixed frozen sections (no antigen retrieval needed)
Positive Control: Kidney, skin, tongue, ovary
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