HIF-1 alpha Transcription Factor Assay 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.
Functional Studies more details
Reacts with: Mouse, Rat, Human; Predicted to work with: all Mammals
Components: 96 tests; 96-Well Plate Cover: 1 unit; Polysorbate 20: 1 vial; Transcription Factor Antibody Binding Buffer (10X): 1 x 3ml; Transcription Factor Binding Assay Buffer (4X): 1 x 3ml; Transcription Factor Developing Solution: 1 x 12ml; Transcription Factor Goat Anti-Rabbit HRP Conjugate: 1 x 100µl; Transcription Factor HIF-1 alpha 96-Well Strip Plate: 1 unit; Transcription Factor HIF-1 alpha Competitor dsDNA: 1 vial; Transcription Factor HIF-1 alpha Positive Control: 1 vial; Transcription Factor HIF-1 alpha Primary Antibody: 1 vial; Transcription Factor Reagent A: 1 x 120µl; Transcription Factor Stop Solution: 1 x 12ml; Wash Buffer Concentrate (400X): 1 x 5ml
Compatible Sample Types:
Cell culture extracts, Cell Lysate