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

Name: HIF-2 alpha IHC Antibody

Clone: Rabbit polyclonal

Supplier: IHC World

Catalog Number: IW-PA1129

Dilution: Ready to Use

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: