Laminin Antibody Staining Protocol for Immunohistochemistry


Description: The extracellular matrix (ECM) is a complex three-dimensional scaffold which surrounds and separates most tissue compartments, determines tissue architecture and dynamically interacts with its surrounding cells. The basement membrane is a part of the ECM and is composed of collagens, proteoglycans, elastin and several glycoproteins of which laminin is the most abundant. Laminins are a family of multifunctional basement membrane glycoproteins with many isoforms composed of variant alpha, beta, and gamma chains. The laminins consist of at least 11 polypeptides (5 alpha chains, 3 beta chains, and 3 gamma chains) specific to basement membranes.


Primary Antibody

Name: Laminin Antibody

Clone: 4C7, Mouse anti-Human

Supplier: Dako

Catalog Number: M0638

Dilution: 1:20 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: Cytoplasmic/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, muscle, placenta
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