Key features and details
- Assay type: Semi-quantitative
- Detection method: Colorimetric
- Platform: Microplate reader
- Sample type: Adherent cells, Nuclear Extracts, Suspension cells
Product namePPAR (alpha, delta, gamma) Transcription Factor Assay Kit
Sample typeAdherent cells, Suspension cells, Nuclear Extracts
Species reactivityReacts with: Human
Predicted to work with: Mammals
Abcam's PPAR Transcription Factor Assay Kit (ab133113) is a non-radioactive, sensitive method for detecting specific transcription factor DNA binding activity in nuclear extracts and whole cell lysates.
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 PPAR response element is immobilized onto the bottom of wells of a 96-well plate. PPARs contained in a nuclear extract, bind specifically to the PPAR response element. PPAR alpha, delta, or gamma are detected by addition of specific primary antibodies directed against the individual PPARs. A secondary antibody conjugated to HRP is added to provide a sensitive colorimetric readout at 450 nm. ab133113 comes with a single plate that measures all three isoforms of PPAR alpha, delta, and gamma. There are enough reagents for one-third of a plate for each isoform.
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. PPARs play pivotal roles in the regulation of lipid metabolism and homeostasis and are important indirect and direct regulators of cellular insulin sensitivity.
Storage instructionsPlease refer to protocols.
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 Complete PPARα Positive Control 1 vial Transcription Factor Complete PPARα Primary Antibody 1 vial Transcription Factor Complete PPARδ Positive Control 1 vial Transcription Factor Complete PPARδ Primary Antibody 1 vial Transcription Factor Complete PPARγ Positive Control 1 vial Transcription Factor Complete PPARγ Primary Antibody 1 vial 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 Reagent A 1 x 120µl Transcription Factor Stop Solution 1 x 12ml Wash Buffer Concentrate (400X) 1 x 5ml
- Pathways and Processes
- Metabolic signaling pathways
- Lipid and lipoprotein metabolism
- Fatty acids
Cellular localizationCytoplasm, nucleus.
- Indomethacin, Non-selective COX inhibitor (ab120719)
- Rosiglitazone, PPARgamma agonist (ab120762)
- Pioglitazone hydrochloride, PPAR-gamma agonist (ab120794)
- Telmisartan, angiotensin II (AT1) antagonist (ab120831)
- Clofibric acid, PPARalpha agonist (ab120833)
- GW 9662, PPARgamma antagonist (ab141125)
- Ciglitazone, PPARgamma agonist (ab141139)
- MK886, Lipoxygenase inhibitor (ab141140)
- 15-Deoxy-Delta12,14-prostaglandin J2, Selective PPARgamma agonist (ab141717)
- Nuclear Extraction Kit (ab221978)
Increasing amounts of positive control (total lysate) are assayed for PPAR gamma DNA-binding activity using ab133113.
Increasing amounts of positive control (total lysate) are assayed for PPAR delta DNA-binding activity using ab133113.
Increasing amounts of positive control (total lysate) are assayed for PPAR alpha DNA-binding activity using ab133113.
ab133113 has been referenced in 3 publications.
- Kasonga A et al. Activation of PPARs Modulates Signalling Pathways and Expression of Regulatory Genes in Osteoclasts Derived from Human CD14+ Monocytes. Int J Mol Sci 20:N/A (2019). PubMed: 30979019
- Zhang Q et al. The interleukin-4/PPAR? signaling axis promotes oligodendrocyte differentiation and remyelination after brain injury. PLoS Biol 17:e3000330 (2019). PubMed: 31226122
- Meex RC et al. ATGL-mediated triglyceride turnover and the regulation of mitochondrial capacity in skeletal muscle. Am J Physiol Endocrinol Metab 308:E960-70 (2015). Functional Studies . PubMed: 25852007