HIF-1alpha Transcription Factor Activity Assay

The HIF-1alpha Transcription Factor Activity Assay from Assay Genie is a DNA-binding immunoassay that quantifies the DNA-binding activity of activated HIF-1alpha in nuclear extracts. Unlike assays that measure total HIF-1alpha protein, this kit uses a microplate pre-coated with a consensus hypoxia response element (HRE) oligonucleotide; activated HIF-1alpha in the sample binds to the immobilized HRE and is detected with a HIF-1alpha-specific primary antibody and an HRP-conjugated secondary reagent, delivering a functional readout of transcription factor activation under hypoxic versus normoxic conditions. Validated for human, mouse and rat reactivity, the assay is widely used in hypoxia, angiogenesis, ischemia-reperfusion and tumor metabolism research, where it complements Western blot by distinguishing transcriptionally active HIF-1alpha from total protein levels. Each kit provides reagents for 96 Assays, internal positive and negative controls, wash buffer and a detailed protocol. Antibody components should be refrigerated; protect from light and limit repeated freeze-thaw cycles. A typical sample volume of 10 to 20 uL nuclear lysate per well is sufficient for accurate quantification.