Human HIF-1 alpha ELISA Kit, Fluorescent is a single-wash 90-min Simplestep used to quantify Human HIF-1 alpha with a sensitivity of 12 pg/ml. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.
- Fluorescent Sandwich ELISA - 530/570/590 nm readout : works on any standard plate reader
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- Frontiers in chemistry 10:8906752022Phthalimide Analogs Enhance Genotoxicity of Cyclophosphamide and Inhibit Its Associated Hypoxia.Applications:Unspecified applicationReactive species:Unspecified reactive speciesAmira M Gamal-Eldeen,Hussein S Agwa,Magdy A-H Zahran,Bassem M Raafat,Sherien M El-Daly,Hamsa J Banjer,Mazen M Almehmadi,Afaf Alharthi,Nahed M Hawsawi,Fayez Althobaiti,Mona A M Abo-ZeidPubMed 35518717
- Frontiers in pharmacology 13:8608982022Perftoran Inhibits Hypoxia-Associated Resistance in Lung Cancer Cells to Carboplatin.Applications:Unspecified applicationReactive species:Unspecified reactive speciesAmira M Gamal-Eldeen,Amani A Alrehaili,Afaf Alharthi,Bassem M RaafatPubMed 35401227
- Frontiers in nutrition 9:8547802022Anti-hypoxic Effect of Polysaccharide Extract of Brown Seaweed in Tongue Squamous Cell Carcinoma.Applications:Unspecified applicationReactive species:Unspecified reactive speciesAmira M Gamal-Eldeen,Bassem M Raafat,Amani A Alrehaili,Sherien M El-Daly,Nahed Hawsawi,Hamsa J Banjer,Eman M Raafat,Mazen M AlmehmadiPubMed 35399691
- Frontiers in pharmacology 13:8441042022Effect of Combined Perftoran and Indocyanine Green-Photodynamic Therapy on HypoxamiRs and OncomiRs in Lung Cancer Cells.Applications:Unspecified applicationReactive species:Unspecified reactive speciesAmira M Gamal-Eldeen,Amani A Alrehaili,Afaf Alharthi,Bassem M RaafatPubMed 35370727
Key facts
- Detection method
- Fluorescent
- Sample types
- Cell culture extracts
- Assay type
- Sandwich (quantitative)
- Reactive species
- Human
- Range
- 0.03 - 120 ng/mL
- Assay time
- 1h 30m
- Sensitivity
- = 12 pg/mL
Reactivity data
| Application | Reactivity | Dilution info | Notes |
|---|---|---|---|
Application sELISA | Reactivity Reacts | Dilution info - | Notes - |
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Target data
Function
The protein expressed by the gene HIF1A functions as a master transcriptional regulator of the adaptive response to hypoxia, activating the transcription of over 40 genes under hypoxic conditions, including erythropoietin, glucose transporters, glycolytic enzymes, vascular endothelial growth factor, HILPDA, and others. These genes' protein products enhance oxygen delivery or facilitate metabolic adaptation to hypoxia. HIF1A is crucial for embryonic vascularization, tumor angiogenesis, and ischemic disease pathophysiology. Its activation requires transcriptional coactivators like CREBBP and EP300, with activity enhanced by interactions with NCOA1 and/or NCOA2. Interaction with redox regulatory protein APEX1 activates CTAD and enhances activation by NCOA1 and CREBBP. Additionally, HIF1A is involved in axonal distribution and mitochondrial transport in neurons during hypoxia. In the context of microbial infection, specifically human coronavirus SARS-CoV-2, HIF1A is necessary for glycolysis induction in monocytes, leading to a proinflammatory state, inducing expression of ACE2, cytokines, and promoting virus replication and monocyte inflammatory response. This supplementary information is collated from multiple sources and compiled automatically.
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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Human HIF-1 alpha ELISA Kit, Fluorescent is a single-wash 90-min Simplestep used to quantify Human HIF-1 alpha with a sensitivity of 12 pg/ml. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.
- Fluorescent Sandwich ELISA - 530/570/590 nm readout : works on any standard plate reader
Key facts
- Detection method
- Fluorescent
- Sample types
- Cell culture extracts
- Assay type
- Sandwich (quantitative)
- Reactive species
- Human
- Range
- 0.03 - 120 ng/mL
- Assay time
- 1h 30m
- Assay Platform
- Pre-coated microplate (12 x 8 well strips)
- Sensitivity
- = 12 pg/mL
Precision
Intra assay
| Sample | n | C.V. |
|---|---|---|
Sample DFO HeLa | n 5 | C.V. 4.3 |
Inter assay
| Sample | n | C.V. |
|---|---|---|
Sample DFO HeLa | n 3 | 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
HIF-1 alpha in vitro CatchPoint® SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of HIF1a protein in human cell extracts.
This CatchPoint SimpleStep ELISA kit has been optimized for Molecular Devices Microplate Readers. Click here for a list of recommended Microplate Readers.
If using a Molecular Devices' plate reader supported by SoftMax® Pro software, a preconfigured protocol for these CatchPoint SimpleStep ELISA Kits is available with all the protocol and analysis settings at www.softmaxpro.org.
The CatchPoint® SimpleStep ELISA® employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. CatchPoint® HRP Development Solution containing the Stoplight Red Substrate is added. During incubation, the substrate is catalyzed by HRP generating a fluorescent product. Signal is generated proportionally to the amount of bound analyte and the intensity is measured in a fluorescence plater reader at 530/570/590 nm Excitation/Cutoff/Emission.
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
Sandwich ELISA - Human HIF-1 alpha ELISA Kit, Fluorescent (ab229433)
Example of human HIF1a standard curve in 1X Cell Extraction Buffer PTR.
The HIF1a standard curve was prepared as described in Section 10. Raw data generated on SpectraMax M4 Multi-Mode Microplate Reader is shown in the table. Background-subtracted data values (mean +/- SD) are graphed.
Sandwich ELISA - Human HIF-1 alpha ELISA Kit, Fluorescent (ab229433)
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. Cells were extracted directly in the culture plate by overlaying culture media with Extraction Buffer PTR (with Extraction Enhancer) such that the final concentration was 1X Extraction Buffer. Extracts were applied to the HIF1 alpha ELISA. Raw data with standard deviation is plotted from triplicate measurements.
Sandwich ELISA - Human HIF-1 alpha ELISA Kit, Fluorescent (ab229433)
Comparison of HIF1 alpha expression in HeLa cell extracts (with and without DFO treatment).
Background subtracted OD450 nm data from three loading concentrations are shown. In the HeLa cell line, DFO treatment is required to detect HIF1 alpha protein.
Sandwich ELISA - Human HIF-1 alpha ELISA Kit, Fluorescent (ab229433)
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.
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