PPAR delta 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 delta is detected by addition of specific primary antibody directed against PPAR delta. A secondary antibody conjugated to HRP is added to provide a sensitive colorimetric readout at 450 nm. ab133~10^6 detects human PPAR delta. This assay does not cross-react with PPAR gamma 1 and PPAR gamma 2 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 delta is the least understood; however, identification of a high-affinity PPAR delta synthetic ligand suggests a potential role for this receptor in lipid metabolism and obesity. PPAR delta is ubiquitously expressed, but it is particularly abundant in tissues such as liver, intestine, kidney, abdominal adipose, and skeletal muscle.
Functional Studies more details
Reacts with: Mouse, 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 PPARδ Positive Control: 1 vial; Transcription Factor PPARδ 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