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Integrin beta1 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
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Name: Integrin beta1 Antibody |
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Clone: 4B7R, Mouse anti-Human |
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Supplier: Santa Cruz Biotechnology |
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Catalog Number: sc-9970 |
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Dilution: 1:50 for paraffin sections and 1:400 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: IHC-TekTM Epitope Retrieval Steamer Set (Cat# IW-1102) |
| Buffer/pH value: IHC-TekTM Epitope Retrieval Solution (Cat# IW-1100) or IHC-TekTM Proteinase K Solution (Cat# IW-1101) |
| Heat/Cool Temperature: 95-100 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 membrane/cytoplasmic |
| Images: Search image |
Additional Information:
| Species Reactivity: Human |
| Fixation: Formalin fixed paraffin sections, or acetone fixed frozen sections. |
| Positive Control: Skin |
| 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: