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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
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Name: Laminin Antibody |
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Clone: LAM-89, Mouse anti-Human |
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Supplier: Vector Labs |
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Catalog Number: VP-L551 |
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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. |
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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 |
References: