Key features and details
- Assay type: Semi-quantitative
- Detection method: Colorimetric
- Platform: Microplate reader
- Sample type: Adherent cells, Nuclear Extracts, Suspension cells
Product namePPAR gamma Transcription Factor Assay Kit
Sample typeAdherent cells, Suspension cells, Nuclear Extracts
Species reactivityReacts with: Mouse, Rat, Human
Predicted to work with: Mammals
PPAR gamma Transcription Factor Assay kit ab133101 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.
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 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
- Pathways and Processes
- Metabolic signaling pathways
- Lipid and lipoprotein metabolism
- Fatty acids
FunctionReceptor that binds peroxisome proliferators such as hypolipidemic drugs and fatty acids. Once activated by a ligand, the receptor binds to a promoter element in the gene for acyl-CoA oxidase and activates its transcription. It therefore controls the peroxisomal beta-oxidation pathway of fatty acids. Key regulator of adipocyte differentiation and glucose homeostasis.
Tissue specificityHighest expression in adipose tissue. Lower in skeletal muscle, spleen, heart and liver. Also detectable in placenta, lung and ovary.
Involvement in diseaseNote=Defects in PPARG can lead to type 2 insulin-resistant diabetes and hyptertension. PPARG mutations may be associated with colon cancer.
Defects in PPARG may be associated with susceptibility to obesity (OBESITY) [MIM:601665]. It is a condition characterized by an increase of body weight beyond the limitation of skeletal and physical requirements, as the result of excessive accumulation of body fat.
Defects in PPARG are the cause of familial partial lipodystrophy type 3 (FPLD3) [MIM:604367]. Familial partial lipodystrophies (FPLD) are a heterogeneous group of genetic disorders characterized by marked loss of subcutaneous (sc) fat from the extremities. Affected individuals show an increased preponderance of insulin resistance, diabetes mellitus and dyslipidemia.
Genetic variations in PPARG can be associated with susceptibility to glioma type 1 (GLM1) [MIM:137800]. Gliomas are central nervous system neoplasms derived from glial cells and comprise astrocytomas, glioblastoma multiforme, oligodendrogliomas, and ependymomas. Note=Polymorphic PPARG alleles have been found to be significantly over-represented among a cohort of American patients with sporadic glioblastoma multiforme suggesting a possible contribution to disease susceptibility.
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)
- 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)
3T3-L1 cells were differentiated to adipocytes with 1 uM dexamethasone (ab120743), 1 ug x mL-1 insulin (ab123768) and 0.5 mM IBMX (ab120840). From the third day, the cells were grown in normal medium with the addition of only 1 ug x mL-1 insulin. From day six, the cells were cultured in normal medium for an additional two days. 40 uL of nuclear extracts were tested in duplicates (+/- SD).
Different volumes of positive control with inhibitor (duplicates, +/-SD).
Different volumes of positive control (duplicates, +/-SD).
Panel A: Increasing amounts of positive control (total lysate) are assayed for PPAR gamma DNA-binding activity using ab133101.
Panel B: PPAR gamma 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.
ab133101 has been referenced in 21 publications.
- Ning Z et al. USP22 regulates lipidome accumulation by stabilizing PPAR? in hepatocellular carcinoma. Nat Commun 13:2187 (2022). PubMed: 35449157
- Comas F et al. Activation of Endogenous H2S Biosynthesis or Supplementation with Exogenous H2S Enhances Adipose Tissue Adipogenesis and Preserves Adipocyte Physiology in Humans. Antioxid Redox Signal 35:319-340 (2021). PubMed: 33554726
- Wagner G et al. LMO3 reprograms visceral adipocyte metabolism during obesity. J Mol Med (Berl) 99:1151-1171 (2021). PubMed: 34018016
- Xu C et al. Pharmacological Activation of RXR-α Promotes Hematoma Absorption via a PPAR-γ-dependent Pathway After Intracerebral Hemorrhage. Neurosci Bull 37:1412-1426 (2021). PubMed: 34142331
- Jiang Y et al. FGF21 Protects against Aggravated Blood-Brain Barrier Disruption after Ischemic Focal Stroke in Diabetic db/db Male Mice via Cerebrovascular PPAR? Activation. Int J Mol Sci 21:N/A (2020). PubMed: 32012810