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Fibronectin Antibody Staining Protocol for Immunohistochemistry
Description: Fibronectin is a glycoprotein that has functions associated with cell adhesion and migration. It is found primarily in basement membranes and is a key extracellular matrix component. This antibody will help detect fibronectin in normal and uncharacterized tissues.
Primary Antibody
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Name: Fibronectin Antibody |
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Clone: 568, Mouse anti-Human |
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Supplier: Vector Labs |
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Catalog Number: VP-F705 |
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Dilution: 1:100 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 minutes/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: Extracellular connective tissue matrix, basement membranes |
| Images: Search image |
Additional Information:
| Tissue Type: Skin, kidney |
| Fixation: Formalin fixed paraffin sections, or acetone fixed frozen sections, or cell smears. |
| Positive Control: Normal kidey (extracellular labeling of connective tissue matrix), basement membrane of skin, blood vessels, smooth muscles, ganglia and muscle fibers. |
| Negative Control: Omit primary antibody, isotype control or 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 |
Notes:
No need to perform antigen retrieval on frozen sections |
References: