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HIF-2 alpha Antibody Staining Protocol for Immunohistochemistry
Description: HIF-2 alpha is also designated EPAS1 whose gene is mapped to 2p21-p16. The predicted mouse protein is 88% identical to human EPAS1. The human EPAS1 gene contains 15 exons and spans at least 120 kb. The positions of the introns within the genomic region encoding the N-terminal bHLH-PAS domains of EPAS1 and AHR are similar, suggesting that the 5-prime ends of the 2 genes may have arisen from a gene duplication event1. Moreover, the predicted protein shares 48% sequence identity with HIF1-alpha, a bHLH-PAS transcription factor that induces EPO gene expression in cultured cells in response to hypoxia. Like HIF1A, EPAS1 binds to and activates transcription from the HIF1A response element derived from the 3-prime flanking region of the EPO gene. EPAS1 is predominantly expressed in highly vascularized tissues of adult humans and in endothelial cells of the mouse adult and embryo. Furthermore, EPAS1 may represent an important regulator of vascularization, perhaps involving the regulation of endothelial cell gene expression in response to hypoxia2. HIF2A is expressed at relatively higher levels in villus sections of placenta and in lung samples compared with other tissues examined3. In addition, The variation in EPAS1 influences the relative contribution of aerobic and anaerobic metabolism and hence the maximum sustainable metabolic power for a given event duration4.
Primary Antibody
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Name: HIF-2 alpha IHC Antibody |
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Clone: Rabbit polyclonal |
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Supplier: IHC World |
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Catalog Number: IW-PA1129 |
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Dilution: Ready to Use |
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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) |
| 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: Nuclear |
| Images: Search image |
Additional Information:
| Species Reactivity: Rat |
| Fixation: Formalin fixed paraffin sections, acetone fixed frozen sections or cultured cells. |
| Positive Control: Rat lung, 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: