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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
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Name: Laminin Antibody |
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Clone: 4C7, Mouse anti-Human |
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Supplier: Dako |
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Catalog Number: M0638 |
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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. |
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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: 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 |
References: