Human Hif1a + BNIP3 Hypoxia In Cell ELISA Kit (IR) is Cell-based (quantitative) ELISA for the measurement of Human Hif1a + BNIP3 Hypoxia In Cell ELISA Kit (IR) production in Human in Suspension cells, Adherent cells samples.
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Key facts
- Detection method
- IR
- Sample types
- Suspension cells, Adherent cells
- Assay type
- Cell-based (quantitative)
- Reactive species
- Human
Reactivity data
| Application | Reactivity | Dilution info | Notes |
|---|---|---|---|
Application ELISA | Reactivity Reacts | Dilution info - | Notes - |
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
What's included?
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Human Hif1a + BNIP3 Hypoxia In Cell ELISA Kit (IR) is Cell-based (quantitative) ELISA for the measurement of Human Hif1a + BNIP3 Hypoxia In Cell ELISA Kit (IR) production in Human in Suspension cells, Adherent cells samples.
Key facts
- Detection method
- IR
- Sample types
- Suspension cells, Adherent cells
- Assay type
- Cell-based (quantitative)
- Reactive species
- Human
- Assay Platform
- Microplate
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
ab129733 is an In-Cell ELISA (ICE) assay kit that uses quantitative immunocytochemistry to measure HIF1 alpha and BNIP3 protein levels in cultured cells. Cells are fixed in a microplate and targets of interest are detected with highly specific, well-characterized antibodies. Relative protein levels are quantified using IRDye®-labeled Secondary Antibodies and IR imaging using a LI-COR® Odyssey® or Aerius® system.
Hypoxia and the cellular response to hypoxic environment is a central topic 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 Hypoxia ICE assay kit provides duplexed measurements of the transcription factor HIF1 alpha and the HIF1A responsive protein BNIP3.
Plates are available in our ICE (In-Cell ELISA) Support Pack (In-Cell ELISA (ICE) Support Pack ab111542) which can be bought seperately.
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.
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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6 product images
In-Cell ELISA - Human Hif1a + BNIP3 Hypoxia In Cell ELISA Kit (IR) (ab129733)
Coefficient of variation for the experiment described in Figure 1.
Western blot - Human Hif1a + BNIP3 Hypoxia In Cell ELISA Kit (IR) (ab129733)
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 - Human Hif1a + BNIP3 Hypoxia In Cell ELISA Kit (IR) (ab129733)
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. BNIP3 staining is nearly undetectable in untreated HeLa cells but is induced by DFO treatment. BNIP3 appears to has a mitochondrial staining pattern in the DFO-treated samples.
Immunocytochemistry/ Immunofluorescence - Human Hif1a + BNIP3 Hypoxia In Cell ELISA Kit (IR) (ab129733)
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. HIF1 alpha 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.
In-Cell ELISA - Human Hif1a + BNIP3 Hypoxia In Cell ELISA Kit (IR) (ab129733)
Sample experiment using ab129733 on HeLa cells treated with a titration of DFO. HeLa cells were seeded to an amine coated 96-well microplate and the following day treated with a titration of DFO. After 24h of DFO exposure, the cells were fixed and stained as described in the protocol and the normalized data is presented here +/- SD (as described in the protocol and data analysis sections). BNIP3 levels are increased with DFO concentrations ≥10µM.
In-Cell ELISA - Human Hif1a + BNIP3 Hypoxia In Cell ELISA Kit (IR) (ab129733)
Sample experiment using ab129733 on HeLa cells treated with a titration of DFO. HeLa cells were seeded to an amine coated 96-well microplate and the following day treated with a titration of DFO. After 24h of DFO exposure, the cells were fixed and stained as described in the protocol and the normalized data is presented here +/- SD (as described in the protocol and data analysis sections). HIF1 alpha results show DFO concentrations ≥10µM induce HIF1A protein levels in a dose dependent manner.
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