PPAR gamma 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 gamma is detected by addition of specific primary antibody directed against PPAR gamma. A secondary antibody conjugated to HRP is added to provide a sensitive colorometric readout at 450 nm. PPAR gamma will not crossreact with PPAR delta or PPAR alpha.
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 gamma is primarily expressed in adipose tissue and to a lesser extent in the colon, immune system and the retina. PPAR gamma was first identified as regulator of adipogenesis, but also plays an important role in cellular differentiation, insulin sensitization, atherosclerosis and cancer.
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 PPAR 96-Well Strip Plate: 1 unit; Transcription Factor PPAR Competitor dsDNA: 1 vial; Transcription Factor PPAR gamma Positive Control: 1 vial; Transcription Factor PPAR gamma 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:
Adherent cells, Suspension cells, Nuclear Extracts