PPAR gamma Transcription Factor Assay Kit (ab133101)
Key features and details
- Assay type: Semi-quantitative
- Detection method: Colorimetric
- Platform: Microplate reader
- Sample type: Adherent cells, Nuclear Extracts, Suspension cells
Overview
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Product name
PPAR gamma Transcription Factor Assay Kit
See all PPAR gamma kits -
Detection method
Colorimetric -
Sample type
Adherent cells, Suspension cells, Nuclear Extracts -
Assay type
Semi-quantitative -
Species reactivity
Reacts with: Mouse, Rat, Human
Predicted to work with: Mammals
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Product overview
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.
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Notes
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.
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Platform
Microplate reader
Properties
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Storage instructions
Please 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 -
Research areas
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Function
Receptor 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 specificity
Highest expression in adipose tissue. Lower in skeletal muscle, spleen, heart and liver. Also detectable in placenta, lung and ovary. -
Involvement in disease
Note=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 similarities
Belongs to the nuclear hormone receptor family. NR1 subfamily.
Contains 1 nuclear receptor DNA-binding domain. -
Cellular localization
Nucleus. - Information by UniProt
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Alternative names
- CIMT1
- GLM1
- NR1C3
see all -
Database links
- Entrez Gene: 5468 Human
- Entrez Gene: 19016 Mouse
- Entrez Gene: 25664 Rat
- Omim: 601487 Human
- SwissProt: P37231 Human
- SwissProt: P37238 Mouse
- SwissProt: O88275 Rat
- Unigene: 162646 Human
see all
Images
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Functional Assay: ab133101 PPAR gamma Transcription Factor Assay Kit
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).
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Functional Assay: ab133101 PPAR gamma Transcription Factor Assay KitDifferent volumes of positive control with inhibitor (duplicates, +/-SD).
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Functional Assay: ab133101 PPAR gamma Transcription Factor Assay KitDifferent volumes of positive control (duplicates, +/-SD).
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Functional Studies - PPAR gamma Transcription Factor Assay Kit (ab133101)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.
