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PPAR gamma Transcription Factor Assay Kit (ab133101)

Reviews (3)Q&A (1)References (21)
Functional Assay: ab133101 PPAR gamma Transcription Factor Assay Kit
  • Functional Assay: ab133101 PPAR gamma Transcription Factor Assay Kit
  • Functional Assay: ab133101 PPAR gamma Transcription Factor Assay Kit
  • Functional Studies - 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

  • 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

  • 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.

  • 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.

  • Platform

    Microplate reader

Properties

Images

  • Functional Assay: ab133101 PPAR gamma Transcription Factor Assay Kit
    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).

  • Functional Assay: ab133101 PPAR gamma Transcription Factor Assay Kit
    Functional Assay: ab133101 PPAR gamma Transcription Factor Assay Kit

    Different volumes of positive control with inhibitor (duplicates, +/-SD).

  • Functional Assay: ab133101 PPAR gamma Transcription Factor Assay Kit
    Functional Assay: ab133101 PPAR gamma Transcription Factor Assay Kit

    Different volumes of positive control (duplicates, +/-SD).

  • Functional Studies - PPAR gamma Transcription Factor Assay Kit (ab133101)
    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.

References (21)

Publishing research using ab133101? Please let us know so that we can cite the reference in this datasheet.

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
View all Publications for this product

Customer reviews and Q&As

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1-4 of 4 Abreviews or Q&A

Kit does not work in porcine nuclear extracts

 Below average Excellent 5/5 (Ease of Use)
Abreviews
We ran this procedure twice, with nuclear extracts from two different tissues (adipose tissue and liver) from pigs, and the kit did not detect any PPAR gamma in the samples. The positive control worked well both times, and there was increasing signal with increasing amounts of positive control. Signal was detected with even the lowest amount of positive control used (0.75uL). However, the competitive inhibitor did not seem to reduce signal as much as expected when positive control was used in those wells (when a sample was used in those wells on the second run, no signal was detected). Our nuclear extracts have been used in western blotting of other transcription factors, but not PPAR gamma therefore, we know the extracts themselves are OK, but they may simply all lack PPAR gamma (unlikely). I also did not realize that this kit does not include a standard curve, so the assay would be relatively quantitative at best, if in fact it worked.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Dr. Elizabeth Dupriest

Verified customer

Submitted Jul 01 2022

Good, fast, reliable.

 Excellent Excellent 5/5 (Ease of Use)
Abreviews
The kit comes with a very detailed manual book. The preparation of the reagents is straightforward. Standard protocol with fast results.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Oct 17 2019

Activated PPAR gamma expression in swine brain subjected to TBI and hemorrhagic shock and treated with normal saline, Hextend, or Hextend + Valproic Acid

 Excellent Excellent 5/5 (Ease of Use)
Abreviews
abreview image
ab133101 - PPARy transcription factor assay kit.

This product was used to detect activated PPARy in swine brain that had been subjected to hemorrhagic shock and TBI and treated with normal saline (NS), Hextend (HEX), or Hextend + valproic acid (HEX + VPA). 40mg of brain per sample was homogenized. Nuclear extraction was performed using an Abcam nuclear extraction kit. Protein levels were balanced. The ELISA was performed per the manufacturer's instructions. Samples were incubated overnight at 4 degrees. Colorimetric reaction was brisk and was monitored for progression (per instructions, at 650 nm). Stop solution was added after 30 minutes of agitation at room temperature (control absorbance = 0.5-0.6). The use of competitive dsDNA produced an expected decrease in absorbance (0.3-0.4). All samples were run in duplicates. The average absorbance for each group was as follows:

SHAM 0.55
NS 0.82
HEX 0.76
HEX + VPA 0.73

These findings were not statistically significant and did not support our hypothesis. As such, they will not be published.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Nov 23 2015

Question

Will this kit test Pparg 1 and 2, or only Pparg 1? Is the primary antibody also available separately?

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Answer

The antibody in the assay is capable of detecting both of the isoforms of PPAR gamma, 1 and 2, which are described at the following UniProt entry:

http://www.uniprot.org/uniprot/P37231

We do not sell the antibody separately, but we do have a similar rabbit polyclonal anti-PPARg antibody that is raised against a C terminus sequence of amino acids that is shared by by both isoforms. The catalogue number is ab19481:

Click here (or use the following: https://www.abcam.com/PPAR-gamma-antibody-ab19481.html).

Read More

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