PPAR alpha Transcription Factor Assay Kit is a non-radioactive, sensitive method for detecting specific transcription factor DNA binding activity in nuclear extracts. 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 peroxisome proliferator response element (PPRE) is immobilized onto the bottom of wells of a 96-well plate. PPARs contained in a nuclear extract, bind specifically to the PPRE. PPAR alpha is detected by addition of specific primary antibody directed against PPAR alpha. A secondary antibody conjugated to HRP is added to provide a sensitive colorometric readout at 450 nm. PPAR alpha Transcription Factor Assay will not cross-react with PPAR gamma or PPAR delta.
Peroxisome proliferator-activated receptors (PPARs) are ligand activated nuclear receptors. Three PPAR subtypes have been identified: alpha, delta and gamma. PPARs can be activated by polyunsaturated fatty acids, eicosanoids and various synthtic ligands.PPAR alpha is predominantly expressed in liver, heart and kidneys. Upon activation by ligands such as free fatty acids, lipids and other small molecules, PPAR alpha forms a heterodimeric complex with RXR alpha, and this complex will bind to response elements allowing transcription of target genes involved in lipid metabolism.
Functional Studies more details
Reacts with: Mouse, Rat, Human
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 PPAR 96-Well Strip Plate: 1 unit; Transcription Factor PPAR alpha Positive Control: 1 vial; Transcription Factor PPAR alpha Primary Antibody: 1 vial; Transcription Factor PPAR Competitor dsDNA: 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:
Adherent cells, Suspension cells, Nuclear Extracts