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HIF-1 alpha Transcription Factor Assay (ab133104) is a non-radioactive, sensitive method for detecting specific transcription factor DNA binding activity in nuclear extracts and whole cell lysate.

Is this your first time purchasing this assay kit? Get 20% off one kit by entering code 20OFF-T5A8Q at checkout. T&C’s apply – click here. Offer only valid when purchasing directly from Abcam in Europe, North America, Australia, and Singapore.

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Images

Key facts

Detection method

Colorimetric

Sample types

Cell culture extracts, Cell Lysate

Assay type

Semi-quantitative

Reactive species

Mouse, Rat, Human

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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:9887100, PubMed:30125331). 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:9887100, PubMed:30125331). 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:9887100, PubMed:16543236). 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 proinflammatory 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

What's included?

96 Test
Components
96-Well Plate Cover
1 x 1 Unit
Polysorbate 20
1 x 1 Vial
Transcription Factor Antibody Binding Buffer (10X)
1 x 3 mL
Transcription Factor Binding Assay Buffer (4X)
1 x 3 mL
Transcription Factor Developing Solution
1 x 12 mL
Transcription Factor Goat Anti-Rabbit HRP Conjugate
1 x 100 µL
Transcription Factor HIF-1 alpha 96-Well Strip Plate
1 x 1 Unit
Transcription Factor HIF-1 alpha Competitor dsDNA
1 x 1 Vial
Transcription Factor HIF-1 alpha Positive Control
1 x 1 Vial
Transcription Factor HIF-1 alpha Primary Antibody
1 x 1 Vial
Transcription Factor Reagent A
1 x 120 µL
Transcription Factor Stop Solution
1 x 12 mL
Wash Buffer Concentrate (400X)
1 x 5 mL

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HIF-1 alpha Transcription Factor Assay (ab133104) is a non-radioactive, sensitive method for detecting specific transcription factor DNA binding activity in nuclear extracts and whole cell lysate.

Is this your first time purchasing this assay kit? Get 20% off one kit by entering code 20OFF-T5A8Q at checkout. T&C’s apply – click here. Offer only valid when purchasing directly from Abcam in Europe, North America, Australia, and Singapore.

Alternative names

Key facts

Detection method

Colorimetric

Sample types

Cell culture extracts, Cell Lysate

Assay type

Semi-quantitative

Reactive species

Mouse, Rat, Human

Assay Platform

Microplate reader

Storage

Shipped at conditions

Dry Ice

Appropriate short-term storage conditions

Multi

Appropriate long-term storage conditions

Multi

Storage information

Please refer to protocols

Notes

HIF-1 alpha Transcription Factor Assay (ab133104) is a non-radioactive, sensitive method for detecting specific transcription factor DNA binding activity in nuclear extracts and whole cell lysate.

A 96-well enzyme-linked immunosorbent assay (ELISA) replaces the cumbersome radioactive electrophoretic mobility shift assay (EMSA). A specific double stranded DNA (dsDNA) sequence containing the HIF-1 alpha response element (5'-ACGTG-3') is immobilized to the wells of a 96-well plate. HIF-1 alpha contained in a nuclear extract, binds specifically to the HIF-1 alpha response element. The HIF transcription factor complex is detected by addition of a specific primary antibody directed against HIF-1 alpha. A secondary antibody conjugated to HRP is added to provide a sensitive colorimetric readout at 450 nm.

Supplementary info

Biological 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.

Mechanical 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.

Pathway

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.

Disease

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

  • Functional Studies - HIF-1 alpha Transcription Factor Assay Kit (ab133104), expandable thumbnail

    Functional Studies - HIF-1 alpha Transcription Factor Assay Kit (ab133104)

    HEK293 cells were treated with 1 mM deferoxamine mesylate (DFO) for 1 or 24 hours. 40 micoliters of nuclear lysates (ab113474; corresponding to 4e6 cells) were tested in duplicates (+/- SD).

  • Functional Studies - HIF-1 alpha Transcription Factor Assay Kit (ab133104), expandable thumbnail

    Functional Studies - HIF-1 alpha Transcription Factor Assay Kit (ab133104)

    Titration of positive control with different volumes of inhibitor (TA), background signal subtracted (duplicates; +/- SD).

  • Functional Studies - HIF-1 alpha Transcription Factor Assay Kit (ab133104), expandable thumbnail

    Functional Studies - HIF-1 alpha Transcription Factor Assay Kit (ab133104)

    Titration of positive control, background signal subtracted (duplicates; +/- SD).

  • Functional Studies - HIF-1 alpha Transcription Factor Assay Kit (ab133104), expandable thumbnail

    Functional Studies - HIF-1 alpha Transcription Factor Assay Kit (ab133104)

    Assay of nuclear extract from stimulated HeLa cells (100 µM CoCl2) and non-stimulated HeLa cells.

Downloads

Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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