Product namePPAR alpha Transcription Factor Assay Kit
Sample typeAdherent cells, Suspension cells, Nuclear Extracts
Species reactivityReacts with: Mouse, Rat, Human
Abcam's PPAR alpha Transcription Factor Assay Kit (ab133107) 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.
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 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
- Pathways and Processes
- Metabolic signaling pathways
- Lipid and lipoprotein metabolism
- Fatty acids
FunctionLigand-activated transcription factor. Key regulator of lipid metabolism. Activated by the endogenous ligand 1-palmitoyl-2-oleoyl-sn-glycerol-3-phosphocholine (16:0/18:1-GPC). Activated by oleylethanolamide, a naturally occurring lipid that regulates satiety (By similarity). Receptor for peroxisome proliferators such as hypolipidemic drugs and fatty acids. Regulates the peroxisomal beta-oxidation pathway of fatty acids. Functions as transcription activator for the ACOX1 and P450 genes. Transactivation activity requires heterodimerization with RXRA and is antagonized by NR2C2.
Tissue specificitySkeletal muscle, liver, heart and kidney.
Sequence similaritiesBelongs to the nuclear hormone receptor family. NR1 subfamily.
Contains 1 nuclear receptor DNA-binding domain.
- Information by UniProt
- 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)
sELISA pair antibody
Panel A: Increasing amounts of positive control (total lysate) are assayed for PPAR alpha DNA-binding activity using ab133107.
Panel B: PPAR alpha DNA-binding assays are performed in the presence of competitive dsDNA. The decrease in signal caused by addition of competitive dsDNA confirms the assay specificity.
This product has been referenced in:
- Qi K et al. Tongxinluo attenuates reperfusion injury in diabetic hearts by angiopoietin-like 4-mediated protection of endothelial barrier integrity via PPAR-a pathway. PLoS One 13:e0198403 (2018). Read more (PubMed: 29912977) »
- Ren T et al. The Combination of Blueberry Juice and Probiotics Ameliorate Non-Alcoholic Steatohepatitis (NASH) by Affecting SREBP-1c/PNPLA-3 Pathway via PPAR-a. Nutrients 9:N/A (2017). Read more (PubMed: 28264426) »