|Product Overview:||The changes of environmental oxygen levels induce a number of metabolic changes with rapid and profound consequences for cell physiology. To regulate oxygen homeostasis, the transcription factor hypoxia-inducible factor (HIF) regulates the expression of up to 50 adaptive genes by binding to hypoxia response elements (HRE) in their promoters. HIF consists of two subunits: oxygen-regulated α-subunit (HIF-1α) and constitutively expressed nuclear β-subunit (HIF-1β). In normoxia, HIF-1α is hydroxylated by oxygen-sensitive prolyl hydroxylases (PHDs) at Pro-402 and/or Pro-564 and asparaginyl hydroxylase FIH at Asn-80. Modified HIF-1α is subsequently destined to be degrade or prevented from the recruitment of the transcriptional coactivators p300 and CBP, resulting in low protein levels within cellular nuclear. Due to high availability of oxygen in hypoxia, however, hydroxylase activity decreases, thus enabling HIF-1a to accumulate and translocate to the nucleus where it forms transcriptional complex with the HIF1β/ARNT subunit and binds to HRE sequence to initiate gene expression. The HIF-1α TF-Activity Assay (Transcription Factor-Activity Assay) kit is a non-radioactive transcription factor assay with an ELISA format. It offers an easy, speedy, sensitive and high-throughput method to detect the activation of transcription factors.|
|Applications:||Detecting the HIF-1α in human nuclear extraction and whole lysates.|
|Kit Components:||Microplate; DNA Binding Buffer (5X); Positive Control; Specific Competitor DNA Probe; Non-specific Competitor DNA Probe; Assay Reagent; DTT (300 mM); Wash Buffer Concentrate (20X); Primary Antibody; HRP-conjugated Secondary Antibody; Antibody Diluent Buffer; TMB One-Step Substrate Reagent; Stop Solution|
|Detection method:||Absorbance (450 nm)|
|Features & Benefits:||A non-radioactive transcription factor assay with an ELISA format.
An easy, speedy, sensitive and high-throughput method to detect the activation of transcription factors.