Human HIF-1 alpha ELISA Kit is a single-wash 90-min Simplestep used to quantify Human HIF-1 alpha with a sensitivity of 42 pg/ml. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.
- Colorimetric Sandwich ELISA - 450 nm readout : works on any standard plate reader
- Cited in over 15 citations
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- Toxics 11:2023A Proposed Association between Improving Energy Metabolism of HepG2 Cells by Plant Extracts and Increasing Their Sensitivity to Doxorubicin.Applications:Unspecified applicationReactive species:Unspecified reactive speciesTamer A Al-Shafie,Engy A Mahrous,Mustafa Shukry,Mohammad Y Alshahrani,Samah F Ibrahim,Liana Fericean,Afaf Abdelkader,Mennatallah A AliPubMed 36851057
- Frontiers in nutrition 9:8572562022Carvacrol Arrests the Proliferation of Hypopharyngeal Carcinoma Cells by Suppressing Ornithine Decarboxylase and Hyaluronidase Activities.Applications:Unspecified applicationReactive species:Unspecified reactive speciesKaneez Fatima,Suaib Luqman,Abha MeenaPubMed 35464036
- Iranian journal of pharmaceutical research : IJPR 20:316-3322021Inhibition of HMGB1 Might Enhance the Protective Effect of Taxifolin in Cardiomyocytes via PI3K/AKT Signaling Pathway.Applications:Unspecified applicationReactive species:Unspecified reactive speciesErjun Feng,Jian Wang,Xinwei Wang,Zhenguo Wang,Xiaochu Chen,Xu Zhu,Wenli HouPubMed 34567165
- International journal of stem cells 14:221-2282021Optimal Hypoxic Preconditioning of Human Embryonic Stem Cell-Derived Mesenchymal Stem Cells (hES-MSCs) and Their Characteristics.Applications:Unspecified applicationReactive species:Unspecified reactive speciesSeung Min Lee,Dae Won Jun,Hyeon Tae Kang,Ju Hee Oh,Waqar Khalid Saeed,Sang Bong AhnPubMed 33632987
- Journal of gastroenterology and hepatology 36:1997-20072021Macro-encapsulation of mesenchymal stem cells in acute and chronic liver injury animal models.Applications:Unspecified applicationReactive species:Unspecified reactive speciesHyeon Tae Kang,Kiseok Jang,Dae Won Jun,Eileen L Yoon,Seung Min Lee,Waqar Khalid Saeed,Jin Ho LeePubMed 33554346
- International journal of molecular sciences 21:2020Hypoxia Regulates DPP4 Expression, Proteolytic Inactivation, and Shedding from Ovarian Cancer Cells.Applications:Unspecified applicationReactive species:Unspecified reactive speciesLaura R Moffitt et. al.PubMed 33143089
- Experimental and therapeutic medicine 20:1412020Expression and predictive value of HIF-1α and VEGF in patients with burns following treatment.Applications:Unspecified applicationReactive species:Unspecified reactive speciesZhufeng Liu et. al.PubMed 33093879
- FEBS letters 595:811-8272020Low oxygen tension differentially regulates the expression of placental solute carriers and ABC transporters.Applications:Unspecified applicationReactive species:Unspecified reactive speciesLudwik Gorczyca,Jianyao Du,Kristin M Bircsak,Xia Wen,Anna M Vetrano,Lauren M AleksunesPubMed 32978975
- BioMed research international 2020:15879152020Association of Nuclear Receptor Coactivators with Hypoxia-Inducible Factor-1 in the Serum of Patients with Chronic Kidney Disease.Applications:Unspecified applicationReactive species:Unspecified reactive speciesTianbiao Zhou et. al.PubMed 32884936
- Cancers 12:2020Chemically Induced Hypoxia Enhances miRNA Functions in Breast Cancer.Applications:Unspecified applicationReactive species:Unspecified reactive speciesEmma Gervin et. al.PubMed 32707933
Key facts
- Detection method
- Colorimetric
- Sample types
- Cell culture extracts
- Assay type
- Sandwich (quantitative)
- Reactive species
- Human
- Range
- 0.23 - 15 ng/mL
- Assay time
- 1h 30m
- Sensitivity
- = 42 pg/mL
Reactivity data
| Application | Reactivity | Dilution info | Notes |
|---|---|---|---|
Application sELISA | Reactivity Reacts | Dilution info - | Notes - |
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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).
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?
Recommended products
Human HIF-1 alpha ELISA Kit is a single-wash 90-min Simplestep used to quantify Human HIF-1 alpha with a sensitivity of 42 pg/ml. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.
- Colorimetric Sandwich ELISA - 450 nm readout : works on any standard plate reader
- Cited in over 15 citations
Key facts
- Detection method
- Colorimetric
- Sample types
- Cell culture extracts
- Assay type
- Sandwich (quantitative)
- Reactive species
- Human
- Range
- 0.23 - 15 ng/mL
- Assay time
- 1h 30m
- Assay Platform
- Microplate
- Sensitivity
- = 42 pg/mL
Precision
Intra assay
| Sample | n | mean | SD | C.V. |
|---|---|---|---|---|
Sample HeLa Extract | n 5 | mean - | SD - | C.V. 4.3 |
Inter assay
| Sample | n | mean | SD | C.V. |
|---|---|---|---|---|
Sample HeLa Extract | n 3 | mean - | SD - | C.V. 7 |
Recovery
Sample specific recovery
| Sample type | Average % | Range |
|---|---|---|
Sample type Bovine Serum Albumin | Average % = 97 | Range 93 - 106 % |
Sample type Cell culture media | Average % = 112 | Range 94 - 129 % |
Sample type Fetal Bovine Serum | Average % = 109 | Range 102 - 120 % |
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
Human HIF1a ELISA kit (ab171577) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of HIF1a protein in human cell extracts. It uses our proprietary SimpleStep ELISA® technology. Quantitate human HIF1a with 42 pg/ml sensitivity.
SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:
-Single-wash protocol reduces assay time to 90 minutes or less
-High sensitivity, specificity and reproducibility from superior antibodies
-Fully validated in biological samples
-96-wells plate breakable into 12 x 8 wells strips
A 384-well SimpleStep ELISA® microplate (Pre-coated 384 well Microplate SimpleStep ELISA® ab203359) is available to use as an alternative to the 96-well microplate provided with SimpeStep ELISA® kits.
Hypoxia-inducible factor 1-alpha (HIF1 alpha) is a constitutively expressed transcription factor that is degraded under normal oxygen tensions but stabilized when oxygen is limiting (hypoxia). Under hypoxic conditions, stabilized HIF1 alpha translocates to the nucleus and promotes the transcription of a host of genes that enable the cell to adapt to the lack of oxygen. Aspects of the HIF1 alpha mediated hypoxic response include promotion of angiogenesis and the switch from aerobic respiration to anaerobic glycolysis. Many of the HIF1 alpha responsive genes encode proteins that promote glycolysis and/or inhibit oxidative phosphorylation (known as the Warburg effect). An exciting and developing area of current cancer research is examining how HIF-mediated metabolic reprogramming promotes tumor growth and survival.
In most cases, HIF1 alpha will need to be stabilized to be measured (steady state levels of HIF1 alpha in non-hypoxic environments is exceeding low in most cell lines). This can be achieved by (a) creating a hypoxic environment (e.g. using a hypoxia chamber) or (b) by using chemical treatments that mimic hypoxia (e.g. cobalt chloride or deferoxamine). The sample data in this assay protocol was generated using deferoxamine (DFO). DFO is an iron chelator and disrupts the function the prolyl hydroxylases that degrade HIF1 alpha in normoxia. By disrupting the enzymes that degrade HIF1 alpha, DFO increases the abundance of HIF1 alpha protein.
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
HIF-1 alpha also known as hypoxia-inducible factor 1-alpha is a transcription factor critical in cellular response to low oxygen levels. Its molecular weight usually ranges from 93 to 120 kDa. You can find HIF-1 alpha expressed in tissues throughout the body but its expression significantly increases under hypoxic conditions. Researchers often use the HIF-1a ELISA to measure its expression levels. HIF-1 alpha forms a complex with other proteins to perform its functions effectively.
- Biological function summary
HIF-1 alpha regulates gene expression in response to hypoxic conditions in cells. It forms a complex with HIF-1 beta to activate transcription of various genes involved in energy metabolism angiogenesis and erythropoiesis. HIF-1 alpha enables cells to adapt to reduced oxygen availability allowing for cellular survival and function under stress. It plays an important role in promoting the expression of genes like VEGF and EPO which are important for vascular and red blood cell development respectively.
- Pathways
HIF-1 alpha plays an integral role in the hypoxia signaling pathway and the glycolytic pathway. In the hypoxia signaling pathway HIF-1 alpha partners with VHL (Von Hippel-Lindau) protein that regulates its degradation under normal oxygen conditions. When oxygen levels drop HIF-1 alpha avoids degradation stabilizes and translocates into the nucleus to initiate transcription of hypoxia-responsive genes. The glycolytic pathway involvement highlights its function in adapting energy production under hypoxic conditions through collaboration with enzymes and transporters associated with glycolysis.
- Associated diseases and disorders
HIF-1 alpha has been implicated in cancer and ischemic diseases. Its role in promoting angiogenesis and metabolic adaptation makes it a contributor to tumor growth and survival collaborating with oncogenes such as c-Myc. In ischemic diseases like stroke or myocardial infarction HIF-1 alpha's ability to induce protective responses can mitigate tissue damage through regulation of survival pathways. Understanding these interactions helps in the development of therapeutic strategies targeting HIF-1 alpha in disease contexts.
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4 product images
ELISA - Human HIF-1 alpha ELISA Kit (ab171577)
Example HIF1 alpha standard curve. Background-subtracted data values (mean +/- SD) are graphed.
ELISA - Human HIF-1 alpha ELISA Kit (ab171577)
Titration of HeLa-DFO (Deferoxamine mesylate, Iron chelator ab120727) extract within the working range of the assay.
Titration of HeLa-DFO extract within the working range of the assay. Background subtracted data from duplicate measurements are plotted. To induce HIF1 alpha protein levels, HeLa cells were treated with 500 μM Deferoxamine (DFO) for 24 hours.
ELISA - Human HIF-1 alpha ELISA Kit (ab171577)
Dose-dependent induction of HIF1 alpha in HeLa cells by DFO (Deferoxamine mesylate, Iron chelator ab120727).
Dose-dependent induction of HIF1 alpha in HeLa cells by deferoxamine (DFO). HeLa cells were cultured in 96-well tissue culture plates and were either untreated or exposed to varying dose of DFO for 24 hours. Raw data with standard deviation is plotted from triplicate measurements.
ELISA - Human HIF-1 alpha ELISA Kit (ab171577)
Comparison of HIF1 alpha expression in HeLa cell extracts (with and without DFO treatment) by ELISA (barchart) and western blot (top). Background subtracted OD450 nm data from three loading concentrations are shown. The HIF1 alpha detector antibody was used to blot the same lysates as analyzed by ELISA (40 μg loaded/lane). The GAPDH blot is included to show the relative loads of each lysate. In the HeLa cell line, DFO treatment is required to detect HIF1 alpha protein by both ELISA and western blot.
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