Pparγ

Manufactured by Santa Cruz Biotechnology

Sourced in United States, China, United Kingdom

PPARγ is a nuclear receptor that functions as a ligand-activated transcription factor. It is involved in the regulation of genes related to cell differentiation, development, and metabolism.

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Lab products found in correlation

Close spelling variants of this product

Anti-PPARγ (75 citations) Anti-PPARγ antibody (21 citations) Mouse anti-PPARγ (11 citations) PPARγ (sc-7273) (10 citations) Anti-PPARγ (sc-7273; (7 citations) Anti-PPARγ (H-100, (6 citations) Mouse monoclonal anti‐PPARγ (5 citations) PPARγ siRNA (5 citations) Antibodies against PPARγ (4 citations) PPARγ (H-100) (4 citations)

177 protocols using pparγ

Antibody Characterization for Western Blotting and Immunoprecipitation

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The following antibodies were used in this study for western blots and immunoprecipitation assays: anti-CHIP (rabbit C3B6; Cell Signaling Technology, Danvers, Massachusetts, USA), MYC (sc-40; Santa Cruz Biotechnology, CA, USA), FLAG (mouse F3165; Sigma-Aldrich, St. Louis, MO, USA), FLAG (rabbit; Sigma-Aldrich), HA (mouse sc-7392; Santa Cruz Biotechnology, CA, USA), PPARγ (mouse sc-7273X, Santa Cruz Biotechnology), PPARγ (rabbit sc-7196X; Santa Cruz Biotechnology), aP2 (goat sc-18661; Santa Cruz Biotechnology), C/EBPα(rabbit sc-61X; Santa Cruz Biotechnology), β-actin (A5316; Sigma-Aldrich), MG132 (M-1157, A.G.Scientific, San Diego, CA, USA), and geldanamycin (9843 S, Cell Signaling Technology). Pepstatin A (P5318), aprotinin (A1153), phenylmethanesulfonylfluoride (PMSF; P7626), leupeptin (L2884), dimethyl sulfoxide (DMSO; D8418), N-ethylmaleimide (NEM; E3876), dexamethasone (D1756), 3-isobutyl-1-methylxanthine (IBMX; I5879), and Oil Red O (O0625) were purchased from Sigma-Aldrich (St. Louis, MO, USA), while insulin (11 376 497 001) was purchased from Roche (Mannheim, Germany).

Kim J.H., Shin S., Seo J., Lee E.W., Jeong M., Lee M.S., Han H.J, & Song J. (2017). C-terminus of HSC70-Interacting Protein (CHIP) Inhibits Adipocyte Differentiation via Ubiquitin- and Proteasome-Mediated Degradation of PPARγ. Scientific Reports, 7, 40023.

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Protein Analysis of Liver and Adipose Tissues

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All reagents were purchased from Sigma-Aldrich (Sigma-Aldrich, St. Louis, MA, USA) unless otherwise specified. Liver tissue was homogenized using a tissue homogenizer in RIPA buffer (50 mM Tris HCL pH:8.0, 150 mM NaCl, 0.1% Triton X-100, 0.5% sodium deoxycholate, 0.1% SDS) supplemented with 2 µg/mL aprotinin (Calbiochem), 5 mM sodium fluoride, 5 mM sodium orthovanadate, and protease inhibitor cocktail (FastPrep^®-24, MP Biomedicals). Adipose tissue protein was extracted using an extraction kit (AT-022, Invent biotechnologies, Plymouth, MN, USA). Lysates were stored at −80 °C for future analysis. Protein extracts were separated by SDS-PAGE 12% polyacrylamide gels as previously reported [11 (link)] and probed for p-AKT (Cell Signaling Technology (CS-4060S), AKT (CS-9272), PPARγ (Santa Cruz Biotechnology-7196), AT2R (abcam92445), p-HSL (CS-41265), HSL (abcam45422), p-PKA (CS-5661S or PKA (CS-58425) overnight before incubation with fluorescent secondary antibodies (Li-cor Biosciences) for 1 h at RT. Images were analyzed using Image Studio Lite software (Li-cor Biosciences, Lincoln, NE, USA). All the phosphorylated proteins bands were normalized to their corresponding total protein. Total proteins were normalized to

β

-actin (Sigma-Aldrich A5441).

de Campos Zani S.C., Wang R., Veida-Silva H., Clugston R.D., Yue J.T., Mori M.A., Wu J, & Chan C.B. (2023). An Egg White-Derived Peptide Enhances Systemic Insulin Sensitivity and Modulates Markers of Non-Alcoholic Fatty Liver Disease in Obese, Insulin Resistant Mice. Metabolites, 13(2), 174.

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Exploring Renal Distal Tubule Epithelium Signaling

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A canine renal distal tubular epithelium cell line (MDCK) was purchased from the American Type Culture Collection (Rockville, USA). RSG, GW9662, and PHA665752 were obtained from Sigma-Aldrich (St. Louis, MO, USA). The following primary antibodies were used: p-Met, Smad7, TGF-β1, and PPAR-γ (Santa Cruz, USA); HGF (Abcam, USA); p-PPAR-γ (Ser112) (Bioss, China); c-Met (Proteintech, China); Smad2, Smad3, p-Smad2 3, and Lamin B (Wanleibio, China); and GAPDH (Zhongshan Golden Bridge Bio Co., Ltd., China). The secondary antibodies, including HRP-conjugated AffiniPure goat anti-rabbit IgG and HRP-conjugated AffiniPure goat anti-mouse IgG, were purchased from Zhongshan Golden Bridge Bio Co., Ltd. (Beijing, China). A Nuclear and Cytoplasmic Protein Extraction Kit was acquired from Beyotime Institute of Biotechnology (Shanghai, China).

Liu Y.D., Yu S.L., Wang R., Liu J.N., Jin Y.S., Li Y.F, & An R.H. (2019). Rosiglitazone Suppresses Calcium Oxalate Crystal Binding and Oxalate-Induced Oxidative Stress in Renal Epithelial Cells by Promoting PPAR-γ Activation and Subsequent Regulation of TGF-β1 and HGF Expression. Oxidative Medicine and Cellular Longevity, 2019, 4826525.

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Protein Fractionation and Analysis

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The total protein of lung tissue was isolated with a protein extraction kit (Nanjing KeyGen Biotech Co., Ltd.) according to the manufacturer's protocol. Nuclear and cytoplasmic fractionations were performed using ER Nuclear and Cytoplasmic Extraction Reagents (Thermo Scientific, USA). Protein concentrations were measured with the BCA Protein Assay kit. The samples were subjected to SDS-polyacrylamide gel electrophoresis (PAGE) in 10% polyacrylamide gels. Immunoblotting was performed with antibodies to NF-κB p65 or IκB or PP AR-γ (Santa Cruz Biotechnology) then using HRP-labeled-goat anti-rabbit antibody. Image Lab was used for quantification.

Xia H., Wang J., Sun S., Wang F., Yang Y., Chen L., Sun Z, & Yao S. (2019). Resolvin D1 Alleviates Ventilator-Induced Lung Injury in Mice by Activating PPARγ/NF-κB Signaling Pathway. BioMed Research International, 2019, 6254587.

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Protein Expression Analysis of Troglitazone-Treated Cells

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Protein lysates from troglitazone-treated TKO MEF, HeLa 229 and H460 cells were resolved by 12% SDS-PAGE electrophoresis, transferred to PVDF membrane, and blocked in 5% non-fat dry milk, and 0.1% TBS-Tween 20. Blots were then probed for ASCT2 (Cell Signaling), GLS1 (Abnova), p-c-MYC (T58) (Abnova), p-c-MYC (S62) (Abnova), c-MYC (Cell Signaling), PPARγ (Santa Cruz Biotechnology), PPARα (Santa Cruz Biotechnology), and β-Actin (Pierce). Protein detection was performed using HRP-conjugated secondary antibodies and ECL Select Western Detection Reagent (GE Life Sciences).

Reynolds M.R, & Clem B.F. (2015). Troglitazone suppresses glutamine metabolism through a PPAR-independent mechanism. Biological chemistry, 396(8), 937-947.

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Protein Expression Analysis of Chondrocyte Differentiation

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After harvesting, total proteins were extracted from the cultures. Protein content was assessed by the Pierce BCA assay kit (Biyuntian, China). Total protein (30 µg) was loaded on to 10% SDS polyacrylamide electrophoresis gels and transferred on to a PVDF membrane. The membrane was immunoblotted overnight at 4°C with antibodies against Col2a1 (1:1000, Abcam, U.S.A.), Sox9 (1:200, Sigma–Aldrich), Runx2 (1:500, Sigma–Aldrich), Mmp13 (1:500, Abcam), Pparγ (1:200, Santa Cruz Biotechnology), Pparγ2 (1:500, Elabscience), Col10a1 (1:200, Abcam), Gapdh (1:2000, Transgen), and β-actin (1:4000, Santa Cruz Biotechnology). The membrane was then incubated with secondary anti-mouse or anti-goat IgG antibodies (1:2000, Transgen) conjugated with peroxidase for 60 min at room temperature. The signal was detected by chemiluminescence using the ECL-Plus Detection System (Transgen). Protein semi-quantitation was based on three independent experiments. The densitometric intensities of protein bands were semi-quantitated using Bandscan 5.0 (Glyko Biomedical, U.S.A.) software and values were normalized to those of β-actin or Gapdh for each sample.

Xu J., Lv S., Hou Y., Xu K., Sun D., Zheng Y., Zhang Z., Li X., Li Y, & Chi G. (2018). miR-27b promotes type II collagen expression by targetting peroxisome proliferator-activated receptor-γ2 during rat articular chondrocyte differentiation. Bioscience Reports, 38(1), BSR20171109.

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Protein Expression Analysis in Adipocytes

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Sybr Green-based qPCR was performed in QuantStudio™ 6 Flex Real-Time PCR System. The ribosomal protein 36B4 (Rplp0) was used as a normalization control. For immunoblotting, total cell lysates were prepared in a lysis buffer containing 50 mM Tris (pH = 7.5), 137 mM NaCl, 1 mM EDTA, 10% glycerol, 1% Triton X-100, and freshly added protease inhibitors. Protein samples were separated by SDS-PAGE and transferred onto PVDF membrane. The membrane was blocked with 5% milk in 1× TBST, incubated with primary and secondary antibodies, and visualized using enhanced chemiluminescence. Primary antibodies against UCP1 (Alpha Diagnosis), PPARγ, Flag-tag (Santa Cruz Biotechnology), FABP4 (Cell Signaling Technology), mitochondrial OXPHOS proteins (MitoSciences), and Myc-tag and Tubulin (Sigma–Aldrich) were used.

Mi L., Zhao X.Y., Li S., Yang G, & Lin J.D. (2016). Conserved function of the long noncoding RNA Blnc1 in brown adipocyte differentiation. Molecular Metabolism, 6(1), 101-110.

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Western Blot Analysis of Oxidative Stress Markers

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Proteins were separated and transferred to an Immobilon membrane (Millipore, Bedford, MA) as previously described51 (link). After electrotransfer, the filters were incubated with appropriate polyclonal antibody against 3-nitrotyrosine (Upstate Biotechnology. Lake Placid. NY), iNOS, phosphorylated Rac1, NOX2, UCP2, CHOP, PPARγ, 3-nitrotyrosine (Santa Cruz Biotechnology, Santa Cruz, CA), and β-actin (Sigma-Aldrich, Alcobendas, Spain). Signals were detected using the ECL Western Blotting Detection Reagent (Amersham Ibérica, Madrid, Spain).

García-Ruiz I., Solís-Muñoz P., Fernández-Moreira D., Grau M., Muñoz-Yagüe T, & Solís-Herruzo J.A. (2016). NADPH oxidase is implicated in the pathogenesis of oxidative phosphorylation dysfunction in mice fed a high-fat diet. Scientific Reports, 6, 23664.

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Comprehensive Western Blot Analysis

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Western blot analysis was performed using a sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) electrophoresis system. Protein samples (20 µg) were resuspended in reduced sample buffer, electrophoresed on a 7.5-10% Tris gel with Tris running buffer, blotted onto PVDF membranes, and sequentially probed with primary antibodies against RUNX2 (Cat. no. 12556; 1:1,000; Cell Signaling Technology), OCN (Cat. no. sc-365797; 1:1,000; Santa Cruz Biotechnology, Inc.), eukaryotic translation initiation factor 4E-binding protein 1 (4E/BP1; Thr37/46; Cat. no. 9644; 1:1,000; Cell Signaling Technology), phospho-S6 ribosomal protein (P-S6; S235/S236; Cat. no. sc-293143; 1:1,000), PPARγ (Cat. no. sc-7273; 1:1,000) (both from Santa Cruz Biotechnology, Inc.), C/EBPβ (Cat. no. 3802; 1:2,000), Sca-1 (Cat. no. 9664; 1:4,000), CD29 (Cat. no. 4706; 1:2,000), CD45 (Cat. no. 13917; 1:6,000), CD11b (Cat. no. 14271; 1:3,000) and β-actin (Cat. no. 3700; 1:2,500) (all from Cell Signaling Technology). Horseradish peroxidase-conjugated goat anti-rabbit (Cat. no. SAB4600223; 1:1,000) or anti-mouse (Cat. no. SAB4600004; 1:1,000) antibodies (both from Sigma) were added, and secondary antibodies were detected using enhanced chemiluminescence (ECL Plus; General Electric Healthcare, Milwaukee, WI, USA).

Wang J., Li S.F., Wang T., Sun C.H., Wang L., Huang M.J., Chen J., Zheng S.W., Wang N., Zhang Y.J, & Chen T.Y. (2017). Isopsoralen-mediated suppression of bone marrow adiposity and attenuation of the adipogenic commitment of bone marrow-derived mesenchymal stem cells. International Journal of Molecular Medicine, 39(3), 527-538.

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Ginsenoside Rg1 Modulates Inflammatory Response

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Ginsenoside Rg1 (Formula: C₄₂H₇₂O₁₄, Figure 1) was purchased from Nanjing Guangrun Biotechnology Co., Ltd. (Nanjing, China) with a purity of ≥ 99% as determined by HPLC. The dexamethasone sodium phosphate injections were purchased from the Cisen Pharmaceutical Co. Ltd. (China). Complete Freund's adjuvant (CFA) (containing 10 mg/mL of dry, heat-killed Mycobacterium tuberculosis) was purchased from Chondrex, Inc. (U.S.). Lipopolysaccharides (LPS) were purchased from Sigma-Aldrich Corporation (U.S.). TNF-α and IL-6 ELISA kits were purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). PPAR-γ, IκBα, p-IκBα and NF-κB (p65 and p-p65) antibodies were obtained from Santa Cruz Biotechnology, Inc. (U.S.). The other chemicals were of analytical grade.

Zhang L., Zhu M., Li M., Du Y., Duan S., Huang Y., Lu Y., Zhang J., Wang T, & Fu F. (2017). Ginsenoside Rg1 attenuates adjuvant-induced arthritis in rats via modulation of PPAR-γ/NF-κB signal pathway. Oncotarget, 8(33), 55384-55393.

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