Reduced samples were loaded onto NuPAGE Bis-Tris (Thermo Fisher Scientific) gels in MOPS buffer and separated proteins were transferred onto a PVDF membrane for immunodetection with anti-PPARγ (C26H12, 1:2000: Cell Signaling Technologies, Danvers, MA, USA), HSP90α (GTX109753: Gene Tex, Irvine, CA, USA, 1:2000), HDAC1 (GTX100513: Gene Tex, 1:2000), GAPDH (GTX100118: Gene Tex, 1:1000), H3K27ac (39134: Active Motif, Carlsbad, CA, USA, 1:2000), H3K9ac (39918: Active Motif, 1:2000), H3k27me3 (C36B11: Cell Signaling Technologies, 1:2000), and Histone H3 (D2B12: Cell Signaling Technologies, 1:2000) antibodies as primary antibodies, and the HRP-conjugated goat anti-rabbit antibody (#7074: Cell Signaling Technologies, 1:2000) as the secondary antibody.
Anti pparγ c26h12
Manufactured by Cell Signaling Technology
Sourced in United States
Anti-PPARγ (C26H12) is a laboratory reagent used to detect and quantify the presence of the peroxisome proliferator-activated receptor gamma (PPARγ) protein in biological samples. It is a monoclonal antibody that specifically binds to the PPARγ protein, allowing researchers to measure its expression levels in cells or tissues.
Automatically generated - may contain errors
Lab products found in correlation
Nupage bis tris, by Thermo Fisher Scientific (1 mentions)
Hsp90α, by GeneTex (1 mentions)
Hdac1, by GeneTex (1 mentions)
H3k9ac, by Active Motif (1 mentions)
Hrp conjugated goat anti rabbit antibody, by Cell Signaling Technology (1 mentions)
Histone h3, by Cell Signaling Technology (1 mentions)
H3k27me3, by Cell Signaling Technology (1 mentions)
H3k27ac, by Active Motif (1 mentions)
Gapdh, by GeneTex (1 mentions)
Anti gapdh fl 335, by Santa Cruz Biotechnology (1 mentions)
Protease inhibitor cocktail, by Roche (1 mentions)
Ez magna chip a g chromatin immunoprecipitation kit, by Merck Group (1 mentions)
Anti phospho akt ser473 d9e, by Cell Signaling Technology (1 mentions)
Anti phospho akt thr308 244f9, by Cell Signaling Technology (1 mentions)
Anti c ebpα, by Cell Signaling Technology (1 mentions)
Anti col1a1 72026, by Cell Signaling Technology (1 mentions)
5 protocols using anti pparγ c26h12
1
Immunoblotting for Nuclear Protein Markers
2
Hypothalamic Gene and Protein Expression
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Peripheral tissues were rapidly dissected, snap frozen, and stored at −80 °C until use. Hypothalamic blocks were micro-dissected from 300 μm coronal brain slices. RNA extraction and qPCR was performed as previously described38 (link) (qPCR primers listed in Supplementary Table S2 ). For protein extraction, frozen tissues were homogenised in Tissue-Protein Extraction Reagent (Peirce) containing Protease Inhibitor Cocktail (Roche) and 1 mM PMSF. Protein levels were determined by SDS-PAGE and Western blot analysis using anti-PPARγ (C26H12, Cell Signalling, 1:500) and anti-GAPDH (FL-335, Santa Cruz Biotechnology, 1:1000) antibodies.
3
ChIP-qPCR Analysis of NF-kB and PPAR-γ
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ChIP was performed by following the protocol of the EZ-Magna ChIP A/G Chromatin Immunoprecipitation kit (Sigma-Aldrich). AML-12 cells and BMDMs (1 x 107) treated with 40 μM genipin or vehicle for 48 h were fixed with 1 % PFA and then washed by PBS. The cells were collected and subjected to cellular and nuclear lysis. The whole nuclear lysate was sheared by a sonicator with optimal conditions (7 s pulse on, 10 s pulse off, 15 cycles, 40 % amplitude) to yield 200-700 pb DNA. 10 % of sheared lysate was used for the Input. 50 μL of the sheared lysate (the equivalent of 1 x 106 cells) was subject to immunoprecipitation by overnight incubation of either anti-NF-kB p65/RelA antibody (A19653, Abclonal), anti- PPARγ (C26H12, Cell Signaling) or IgG control. The immunoprecipitated DNA and Input DNA were purified and amplified by RT-PCR with primers sets listed above.
4
Adipogenic Differentiation of Rat MSCs
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Mesenchymal stromal cells (MSCs) from 12-week old WT and Cyp2j4−/− rats were obtained as previously described [32] (link). MSCs cells were allowed to grow in Supplemented MesenCult™ MSC Medium (STEMCELL Technologies, UK) for 5 days on Petri dishes (Nunc, ThermoFisher Scientific, UK). MSCs from WT and Cyp2j4−/− rats were differentiated into mature adipocytes by incubation with an adipogenic induction medium (StemPro®,Gibco, UK) for 14 days.
Antibodies used in western blot were: anti-PPAR-γ (C26 H12, Cell Signaling #2435, 1:1000), anti CEBP-α (Cell Signaling #2295, 1:1000), anti-Phospho-Akt-Ser473 (D9E, Cell Signaling #4060, 1:2000), anti-Phospho-Akt-Thr308 (244F9, Cell Signaling #4056, 1:1000), anti-panAkt (C67E7, Cell Signaling #4691, 1:1000) and anti-β-Actin Antibody (C4, sc-47778, 1:10,000), anti-PPARα (H2, SC-398,394, 1:1000), anti-PPARβ/δ (F-10, SC-74517, 1:1000), anti-FXR (D-3, SC-25309, 1:1000), anti-LXRα (ab2585, 1:1000), and anti-β-Actin Antibody (C4, sc-47778, 1:10,000).
Antibodies used in western blot were: anti-PPAR-γ (C26 H12, Cell Signaling #2435, 1:1000), anti CEBP-α (Cell Signaling #2295, 1:1000), anti-Phospho-Akt-Ser473 (D9E, Cell Signaling #4060, 1:2000), anti-Phospho-Akt-Thr308 (244F9, Cell Signaling #4056, 1:1000), anti-panAkt (C67E7, Cell Signaling #4691, 1:1000) and anti-β-Actin Antibody (C4, sc-47778, 1:10,000), anti-PPARα (H2, SC-398,394, 1:1000), anti-PPARβ/δ (F-10, SC-74517, 1:1000), anti-FXR (D-3, SC-25309, 1:1000), anti-LXRα (ab2585, 1:1000), and anti-β-Actin Antibody (C4, sc-47778, 1:10,000).
5
Antibody-based Protein Expression Analysis
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The antibodies used in the present study were as follows: anti-COL1A1 (72026) and anti-PPARγ (C26H12) from Cell Signaling Technologies (Boston, USA); and anti-ITIH4 (ab180139) from Abcam (Cambridge, UK). Secondary antibodies were either goat Alexa 488 (A-11008) or 594 (A-11012) obtained from ThermoFisher (Oxford, UK). Lipopolysaccharide (LPS, L2630) was purchased from Sigma (Steinheim, Germany).
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