HIF-1 alpha Antibody Staining Protocol for Immunohistochemistry

 

Description: HIF-1α (Hypoxia-inducible factor 1α,HIF1A) is a transcription factor that mediates cellular and systemic homeostatic responses to reduced O2 availability in mammals, including angiogenesis, erythropoiesis and glycolysis.  This gene was mapped to 14q21-q24. HIF-1α transactivate genes required for energy metabolism and tissue perfusion and is necessary for embryonic development and tumor explant growth. HIF-1alpha is over expressed during carcinogenesis, myocardial infarction and wound healing.  It is crucial for the cellular response to hypoxia and is frequently over expressed in human cancers, resulting in the activation of genes essential for cell survival.  HIF-1α regulates the survival and function in the inflammatory microenvironment directly. It is a transcription factor that plays a pivotal role in cellular adaptation to changes in oxygen availability.

 

Primary Antibody

Name: HIF-1 alpha IHC Antibody

Clone: Rabbit polyclonal

Supplier: IHC World

Catalog Number: IW-PA1041

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: Cytoplasmic in normoxia, nuclear in hypoxia.

Images: Search image

Additional Information:

Species Reactivity: Human, mouse, rat, rabbit

Fixation: Formalin fixed paraffin sections

Positive Control: Skin, hypoxia induced tissues, mammary cancer tissues.

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

 

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