Hypoxia and the cellular response to hypoxic environment are central topics in studies of metabolism, cancer progression and development and stem cells.
Images
Key facts
- Detection method
- Fluorescent
- Assay Platform
- Flow cytometer
- Sample type
- Suspension cells, Adherent cells
- Assay type
- Sandwich
- Reactive species
- Human
Target data
Function
Functions as a master transcriptional regulator of the adaptive response to hypoxia (PubMed:11292861, PubMed:11566883, PubMed:15465032, PubMed:16973622, PubMed:17610843, PubMed:18658046, PubMed:20624928, PubMed:22009797, PubMed:30125331, PubMed:9887100). Under hypoxic conditions, activates the transcription of over 40 genes, including erythropoietin, glucose transporters, glycolytic enzymes, vascular endothelial growth factor, HILPDA, and other genes whose protein products increase oxygen delivery or facilitate metabolic adaptation to hypoxia (PubMed:11292861, PubMed:11566883, PubMed:15465032, PubMed:16973622, PubMed:17610843, PubMed:20624928, PubMed:22009797, PubMed:30125331, PubMed:9887100). Plays an essential role in embryonic vascularization, tumor angiogenesis and pathophysiology of ischemic disease (PubMed:22009797). Heterodimerizes with ARNT; heterodimer binds to core DNA sequence 5'-TACGTG-3' within the hypoxia response element (HRE) of target gene promoters (By similarity). Activation requires recruitment of transcriptional coactivators such as CREBBP and EP300 (PubMed:16543236, PubMed:9887100). Activity is enhanced by interaction with NCOA1 and/or NCOA2 (PubMed:10594042). Interaction with redox regulatory protein APEX1 seems to activate CTAD and potentiates activation by NCOA1 and CREBBP (PubMed:10202154, PubMed:10594042). Involved in the axonal distribution and transport of mitochondria in neurons during hypoxia (PubMed:19528298). (Microbial infection) Upon infection by human coronavirus SARS-CoV-2, is required for induction of glycolysis in monocytes and the consequent pro-inflammatory state (PubMed:32697943). In monocytes, induces expression of ACE2 and cytokines such as IL1B, TNF, IL6, and interferons (PubMed:32697943). Promotes human coronavirus SARS-CoV-2 replication and monocyte inflammatory response (PubMed:32697943).
Additional Targets
BNIP3
Alternative names
BHLHE78, MOP1, PASD8, HIF1A, Hypoxia-inducible factor 1-alpha, HIF-1-alpha, HIF1-alpha, ARNT-interacting protein, Basic-helix-loop-helix-PAS protein MOP1, Class E basic helix-loop-helix protein 78, Member of PAS protein 1, PAS domain-containing protein 8, bHLHe78
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Hypoxia and the cellular response to hypoxic environment are central topics in studies of metabolism, cancer progression and development and stem cells.
Key facts
- Detection method
- Fluorescent
- Assay Platform
- Flow cytometer
- Sample type
- Suspension cells, Adherent cells
- Assay type
- Sandwich
- Reactive species
- Human
Storage
- Shipped at conditions
- Blue Ice
- Appropriate short-term storage conditions
- +4°C
- Appropriate long-term storage conditions
- +4°C
- Storage information
- +4°C
Notes
Hypoxia and the cellular response to hypoxic environment are central topics in studies of metabolism, cancer progression and development and stem cells. A key player is the transcription factor HIF1 alpha (hypoxia inducible factor 1 alpha) which is stabilized at the protein level in response to decreased oxygen tension. HIF1 alpha then promotes transcription of a number of factors that alters cellular physiology. This Hypoxic Response Human Flow Cytometry Kit provides duplexed measurements of the transcription factor HIF1 alpha and the HIF1 alpha responsive protein BNIP3.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
HIF1a also known as Hypoxia-Inducible Factor 1-alpha is a transcription factor with a mass of approximately 92 kDa. It functions mainly in response to low oxygen levels (hypoxia) within cells. HIF1a exists in the cytoplasm under normal oxygen conditions and translocates to the nucleus during hypoxia. It is expressed in various tissues and cells particularly those located in areas with fluctuating oxygen levels such as skeletal muscle and heart tissues. The protein also shows up in less oxygenated environments like tumors due to its role in adapting to and surviving low oxygen conditions.
- Biological function summary
HIF1a plays a major role in oxygen homeostasis by regulating the expression of genes involved in important processes such as angiogenesis metabolism and cell survival. HIF1a forms a heterodimeric complex with HIF1b enabling DNA binding to hypoxia response elements of target genes. Additionally BNIP3 an apoptosis-inducing protein regulated by HIF1a under hypoxic conditions promotes cell death and autophagy. The interplay between HIF1a and BNIP3 contributes to maintaining cellular energy balance and responding to stress.
- Pathways
HIF1a is an important player in the hypoxia response pathway which involves numerous transcriptional targets that control oxygen-sensitive processes. It tightly associates with the VEGF (vascular endothelial growth factor) pathway to promote angiogenesis. HIF1a also links with other proteins like PHDs (prolyl hydroxylases) which play a part in the oxygen-sensing mechanism by regulating the degradation of HIF1a under normoxic conditions. Through these pathways HIF1a ensures cells adapt to and survive fluctuations in oxygen availability.
- Associated diseases and disorders
HIF1a has significant roles in cancer and cardiovascular diseases due to its involvement in promoting cell survival angiogenesis and metabolic reprogramming. Elevated levels of HIF1a are commonly found in many tumors where it drives tumor progression through enhanced glucose metabolism and blood vessel formation. In cardiac ischemia HIF1a activation aids tissue adaptation to oxygen deprivation and may protect against heart failure. Its role in both cancer and cardiovascular disease often links with BNIP3 where it contributes to disease pathology by modulating programmed cell death and adaptation mechanisms.
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5 product images
Flow Cytometry - HIF1a + BNIP3 Hypoxia Response Human Flow Cytometry Kit (ab126585)
Figure 2. Sample experiment using ab126585 on HeLa cells treated with a titration of DFO: BNIP3 readout. HeLa cells were cultured in standard tissue culture plates and treated with a titration of DFO. After 24 hours of DFO exposure, the cells were harvested, fixed and stained as described in the protocol. (A) Flow cytometry histogram showing mean fluorescent intensity of BNIP3 staining for untreated (Vehicle) and DFO treated samples. In this experiment anti-mouse-DyLight®488 (Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed ab96879, 1:1000) was used as the secondary antibody and the signal was collected in FL1. (B) Plot showing fold induction of BNIP3 levels (relative to untreated cells) as a function of DFO concentration (blue line). The gray dotted line demarks 1 (the untreated level). DFO concentrations =10μM induce BNIP3 protein levels in a dose dependent manner.
Western blot - HIF1a + BNIP3 Hypoxia Response Human Flow Cytometry Kit (ab126585)
Figure 5. Antibody specificity demonstrated by Western Blot. Primary antibodies used in this assay kit were validated by Western Blot using HeLa cell lysates that had been treated with a dose titration of DFO as indicated. (A) The HIF1 alpha band (indicated by arrow) is absent in untreated cells and induced by DFO. (B) Similarly, BNIP3 levels are increased by DFO treatment in a dose-dependent manner.
Immunocytochemistry/ Immunofluorescence - HIF1a + BNIP3 Hypoxia Response Human Flow Cytometry Kit (ab126585)
Figure 4. Antibody specificity demonstrated by immunocytochemistry. Primary antibodies used in this assay kit were validated by staining HeLa cells +/- treatment with 1mM DFO (24h) and imaged by fluorescent microscopy. Staining of the BNIP3 antibody from this kit is nearly undetectable in untreated HeLa cells but is induced by DFO treatment. BNIP3 appears to have a mitochondrial staining pattern in the DFO-treated samples.
Flow Cytometry - HIF1a + BNIP3 Hypoxia Response Human Flow Cytometry Kit (ab126585)
Figure 1. Sample experiment using ab126585 on HeLa cells treated with a titration of DFO: HIF1 alpha readout. HeLa cells were cultured in standard tissue culture plates and treated with a titration of DFO. After 24 hours of DFO exposure, the cells were harvested, fixed and stained as described in the protocol. (A) Flow cytometry histogram showing mean fluorescent intensity of HIF1 alpha staining for untreated (Vehicle) and DFO treated samples. In this experiment anti-rabbit-DyLight®650 (Goat Anti-Rabbit IgG H&L (DyLight® 650) preadsorbed ab96902, 1:2000) was used as the secondary antibody and the signal was collected in FL4. (B) Plot showing fold induction of HIF1 alpha levels (relative to untreated cells) as a function of DFO concentration (red line). The gray dotted line demarks 1 (the untreated level). DFO concentrations =10µM induce HIF1A protein levels in a dose dependent manner.
Immunocytochemistry/ Immunofluorescence - HIF1a + BNIP3 Hypoxia Response Human Flow Cytometry Kit (ab126585)
Figure 3. Antibody specificity demonstrated by immunocytochemistry. Primary antibodies used in this assay kit were validated by staining HeLa cells +/- treatment with 1mM DFO (24h) and imaged by fluorescent microscopy. Staining is absent in untreated cells and induced by DFO treatment. HIF1 alpha localizes to the nucleus (as seen by co-localization with the DNA stain DAPI) as expected.
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