Laminin Antibody Staining Protocol for Immunohistochemistry

 

Description: Laminin is a large extracellular matrix glycoprotein found in basement membranes of epithelia, surrounding blood vessels and nerves in established tissues. It interacts with cell surface receptors and has roles in cell migration during embryonic development and tissue organization. This antibody will help detect laminin and may be useful as a marker for basement membrane integrity.

 

Primary Antibody

Name: Laminin Antibody

Clone: LAM-89, Mouse anti-Human

Supplier: Vector Labs

Catalog Number: VP-L551

Dilution: 1:50 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

Counterstain
Reagent: Mayer's Hematoxylin
Staining Time: 30 seconds

Results:
Staining Pattern: Basement membranes of blood vessels, smooth muscle, ganglia and muscle fibers.
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, skeletal muscle, small intestine.
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

 

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