Dulbecco minimum essential medium (DMEM) high glucose, fetal bovine serum (FBS), L-glutamine, penicillin and streptomycin, were purchased from Invitrogen Life Technologies (Grand Island, NY, USA). Adipogenic induction medium (AIM) was provided by ScienCell (Carlsbad, CA, USA). All the primers used in this study were synthesized by Invitrogen Life Technologies. Oil Red O, Hoescht 33258, Dexamethasone, β-Glycerophosphate and ascorbic acid were provided by Sigma Alrich (Sigma Alrich, St Louis, MO, USA). Mouse anti-PPARγ antibody was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA), Rabbit anti-adipocyte protein2 (AP2) antibody was provided by Abcam (MA, USA), Rabbit anti-lipoprotein lipase (LPL) and rabbit anti-GAPDH antibody were obtained from Cell Signaling Technology (Beverly, MA, USA). CellTiter 96® AQueous One Solution Cell Proliferation Assay was obtained from Promega (Madison, WI, USA). BCIP/NBT ALP Color Development Kit was purchased from Beyotime (Shanghai, China). Enhanced chemiluminescence (ECL) reagent was provided by Pierce Technology (Pierce, USA).
Mouse anti pparγ antibody
Manufactured by Santa Cruz Biotechnology
Sourced in United States
The Mouse anti-PPARγ antibody is a primary antibody that specifically binds to and recognizes the Peroxisome Proliferator-Activated Receptor Gamma (PPARγ) protein. PPARγ is a nuclear receptor that plays a key role in the regulation of adipogenesis, glucose and lipid metabolism.
Automatically generated - may contain errors
Lab products found in correlation
Celltiter 96 aqueous one solution cell proliferation assay, by Promega (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Dexamethasone (dex), by Merck Group (1 mentions)
Hoescht 33258, by Merck Group (1 mentions)
Bcip nbt alp color development kit, by Beyotime (1 mentions)
β glycerophosphate, by Merck Group (1 mentions)
Rabbit anti gapdh antibody, by Cell Signaling Technology (1 mentions)
Oil red o, by Merck Group (1 mentions)
Ascorbic acid, by Merck Group (1 mentions)
Chemidoc xrs imaging system, by Bio-Rad (1 mentions)
Ip lysis buffer, by Cell Signaling Technology (1 mentions)
Protein a g agarose, by Abcam (1 mentions)
4 protocols using mouse anti pparγ antibody
1
Adipogenic Differentiation of Cells
2
Rat Liver Protein Expression Analysis
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Rat liver tissues (0.02g per specimen) were homogenized and subsequently lysed with ice-cold lysis buffer. Next, the samples were collected and centrifuged at 4°C, at a speed of 10000 rpm for 10 min, following which, the supernatant was stored at −80°C until final use. Western blot assays were performed as previously described [7 (link)], and were conducted with a mouse anti-PPAR-γ antibody (Santa Cruz Biotechnology, code sc-1981; Santa Cruz, U.S.A.; diluted 1:500), a rabbit anti-phosphorylated NF-κB p65 (p-NF-κBp65) antibody (Wanlei Biotechnology, code WL02169, China; diluted 1:500), a rabbit anti-cyclooxygenase-2 (COX-2) antibody (Abcam, code AB15191, U.S.A.; diluted 1:4000), and a mouse anti-GAPDH antibody (Protech, code 10494-1-AP, U.S.A.; diluted 1:8000). Quantification of proteins was analyzed using the Bio-Rad ChemiDoc XRS+ imaging system (U.S.A.). In this analysis, we used a random number table to select three rats from each group. All Western blot images were measured three times.
3
Western Blot Analysis of Transcription Factors
4
Protein Interactions in Adipogenesis
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Mouse PKCα, PRDM16, and PGC1α genes were cloned into the pcDNA3.1(-) vector. HEK293T cells were co-transfected with these genes and PPARγ2. After 24 h, the cells were lysed in Western blotting and IP lysis buffer (Cell Signaling Technology, 9803 S). Total protein (500 μg − 1 mg) was pre-cleared with 1 μg rabbit IgG (Cell Signaling Technology, #3900) and 20 μL Protein A + G agarose (Abcam, ab193262) for 3 h at 4 °C. Total protein was isolated by centrifugation and 0.5 μg anti-Flag antibody was added. Following an overnight incubation at 4 °C, 30 μL Protein A + G agarose was used to purify the immune complex, following which the agarose was washed 7 times in wash buffer (150 mM NaCl, 1 mM EDTA, 20 mM Tris, 1% Triton X-100, pH 8.0). Protein was eluted in 1× SDS-PAGE loading buffer at 100 °C for 10 min. For Co-IP assays in primary adipocytes, mature adipocytes differentiated in vitro were collected and lysed, and 2 μg mouse anti-PPARγ antibody (Santa Cruz, sc-7273) was used for IP.
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