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PPAR gamma Transcription Factor Assay试剂盒(ab133101)

Reviews (3)Q&A (1)References (23)
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

概述

  • 产品名称

    PPAR gamma Transcription Factor Assay试剂盒

  • 检测方法

    Colorimetric

  • 样品类型

    Adherent cells, Suspension cells, Nuclear Extracts

  • 检测类型

    Semi-quantitative

  • 种属反应性

    与反应: Mouse, Rat, Human
    预测可用于: Mammals

  • 产品概述

    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.

  • 说明

    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.

  • 平台

    Microplate reader

性能

图片

  • 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.

文献 (23)

发表研究结果有使用 ab133101？请让我们知道，以便我们可以引用本数据表中的参考文章。

ab133101 被引用在 23 文献中.

  • Ma L  et al. Exercise protects aged mice against coronary endothelial senescence via FUNDC1-dependent mitophagy. Redox Biol 62:102693 (2023). PubMed: 37030149
  • Kim SK  et al. HMG-CoA Reductase Inhibitors Suppress Monosodium Urate-Induced NLRP3 Inflammasome Activation through Peroxisome Proliferator-Activated Receptor-γ Activation in THP-1 Cells. Pharmaceuticals (Basel) 16:N/A (2023). PubMed: 37111279
  • 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
View all Publications for this product

客户评价及客户问答

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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

提交于 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

提交于 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

提交于 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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