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Hypoxia-Inducible Factor 2 alpha (HIF-2-alpha) Antibody Staining Protocol for Immunohistochemistry
Description: Hypoxia contributes significantly to the pathophysiology of major categories of human disease, including myocardial and cerebral ischemia, cancer, pulmonary hypertension, congenital heart disease and chronic obstructive pulmonary disease. HIF-2 alpha is predominantly expressed in highly vascularized tissues of adult humans and endothelial cells of the embryonic and adult mouse, whereas HIF-1alpha functions primarily in extravascular tissues. HIF-2 alpha is also a potent activator of the tie-2 gene, which is known to be selectively expressed in endothelial cells.
Primary Antibody
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Name: HIF-2-alpha Antibody |
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Clone: Rabbit polyclonal |
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Supplier: Novus Biologicals |
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Catalog Number: NB100-122 |
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Dilution: 1:300 - 1:500 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: IHC-TekTM Epitope Retrieval Steamer Set (Cat# IW-1102) |
| Buffer/pH value: IHC-TekTM Epitope Retrieval Solution (Cat# IW-1100) |
| Heat/Cool Temperature: 95 ºC - 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: Nuclear/cytoplasmic |
| Images: Search image |
Additional Information:
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Species Reactivity: Mouse, rat, human |
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Fixation: Formalin fixed paraffin sections |
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Positive Control: Mouse skin |
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Negative Control: Omit primary antibody, isotype control or absorption control |
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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:
Not tested on frozen sections. |
References: