Key features and details
- Assay type: Semi-quantitative
- Detection method: Colorimetric
- Platform: Microplate reader
- Sample type: Adherent cells, Nuclear Extracts, Suspension cells
Product namePPAR delta Transcription Factor Assay Kit
Sample typeAdherent cells, Suspension cells, Nuclear Extracts
Species reactivityReacts with: Mouse, Human
Predicted to work with: Mammals
Abcam's PPAR delta Transcription Factor Assay Kit (ab133106) 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. ab133106 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.
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 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
- Pathways and Processes
- Metabolic signaling pathways
- Lipid and lipoprotein metabolism
- Fatty acids
FunctionLigand-activated transcription factor. Receptor that binds peroxisome proliferators such as hypolipidemic drugs and fatty acids. Has a preference for poly-unsaturated fatty acids, such as gamma-linoleic acid and eicosapentanoic acid. Once activated by a ligand, the receptor binds to promoter elements of target genes. Regulates the peroxisomal beta-oxidation pathway of fatty acids. Functions as transcription activator for the acyl-CoA oxidase gene. Decreases expression of NPC1L1 once activated by a ligand.
Tissue specificityUbiquitous with maximal levels in placenta and skeletal muscle.
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)
- 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 delta DNA-binding activity using ab133106.
Panel B: PPAR delta 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.
ab133106 has been referenced in 2 publications.
- Huang KC et al. Stearoyl-CoA Desaturase-1 Attenuates the High Shear Force Damage Effect on Human MG63 Osteosarcoma Cells. Int J Mol Sci 21:N/A (2020). PubMed: 32630668
- Sharif O et al. The triggering receptor expressed on myeloid cells 2 inhibits complement component 1q effector mechanisms and exerts detrimental effects during pneumococcal pneumonia. PLoS Pathog 10:e1004167 (2014). Functional Studies ; Mouse . PubMed: 24945405