Pparγ

Manufactured by Thermo Fisher Scientific

Sourced in United States

PPARγ is a receptor protein that functions as a transcription factor, regulating the expression of genes involved in cell differentiation and metabolism. It plays a key role in adipocyte differentiation and is important in the regulation of glucose and lipid homeostasis.

Automatically generated - may contain errors

Lab products found in correlation

23 protocols using pparγ

Immunofluorescence Staining of SREBP-1 and PPAR-γ

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Immunofluorescence staining was performed as previously described [16 (link)]. The primary antibodies used were PPAR- γ (Cell Signaling Technology) and SREBP-1 (Santa Cruz Biotechnology, Santa Cruz, CA, USA). The secondary antibodies used were Alexa Fluor 488 and/or Alexa Fluor 594.
The SZ95 cells were fixed with 10% formalin for 15 min at room temperature. The cells were subsequently treated with 0.1% Triton X-100 in PBS for 5 min to permeabilize them. Then, the cells were blocked at room temperature for 30 min and incubated with primary antibody against SREBP-1 and PPAR-γ at 37 °C for 1 h. For double staining, a reaction with Alexa Fluor 488 phalloidin (Thermo Fisher Scientific, Waltham, MA, USA) was performed at room temperature for 1 h. The nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI) for 5 min at 37 °C. The slides were mounted using VECTASHIELD Mounting Medium (VECTOR Laboratories, Burlingame, CA, USA).

Gu H., An H.J., Gwon M.G., Bae S., Zouboulis C.C, & Park K.K. (2022). The Effects of Synthetic SREBP-1 and PPAR-γ Decoy Oligodeoxynucleotide on Acne-like Disease In Vivo and In Vitro via Lipogenic Regulation. Biomolecules, 12(12), 1858.

+ Open protocol

+ Expand

Adipose Tissue Protein Expression Analysis

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

At day one after birth, adipose tissue was collected from the pups that were culled to standardize the litter size. Newborns were decapitated and inguinal (subcutaneous) adipose tissue was pooled from 4 males per litter. At 9 months of age, retroperitoneal (visceral) adipose tissue, which is associated with obesity and insulin resistance^{21 (link),22 (link)} was collected and samples frozen in liquid nitrogen and stored at −80°C till protein analysis. To control for litter effects in the adult studies, one male was studied from each of the litters. Thus when n = 6, it represents one male from each of 6 different litters in Control, IUGR and Mat-OB groups.
Protein was extracted in radioimmuno precipitation assay (RIPA) buffer that contained protease inhibitors (HALT cocktail, Pierce). Supernatant protein concentration was determined by BCA solution (PIERCE, Rockford, IL). Protein expression was determined by Western Blotting, as previously described.^{15 (link)} Antibodies were obtained from Santa Cruz, CA unless otherwise specified and the band density was analyzed as indicated: PPARγ (57 kDa; Thermo Scientific, Rockford, IL), C/EBPα (42 kDa), SREBP1c (125 kDa), FAS (270 kDa), hormone sensitive lipase (84 kDa), SIRT1 (120 kDa), NCoR (270 kDa), SMRT (160 kDa), SRC1 (160 kDa), TIF2 (158 kDa) and β-actin (Sigma, A-5441; 40 kDa).

Desai M., Jellyman J.K., Han G., Lane R.H, & Ross M.G. (2015). Programmed Regulation of Rat Offspring Adipogenic Transcription Factor (PPARγ) by Maternal Nutrition. Journal of developmental origins of health and disease, 6(6), 530-538.

+ Open protocol

+ Expand

Western Blot Analysis of Circadian Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols

Briefly, homogenization buffer was added to tissues which were then homogenized on ice, centrifuged, and the supernatant carefully removed. Basing on Bradford protein quantification, the 50 μg protein lysate was electrophoretically fractionated through a 12% SDS-polyacrylamide gel and electrotransferred onto a polyvinylidene difluoride (PVDF) membrane (Amersham, Piscataway, NJ, USA). The blots were soaked in blocking buffer for one hour at room temperature to block non-specific protein-binding sites. The primary antibodies for BMAL1 (EMD Millipore, Billerica, MA, USA; 1/2500), PPARγ (Thermo Fisher Scientific, Waltham, MA, USA; 1/4000) and β-Actin (EMD Millipore; 1/20000) were diluted in TBST buffer according to the titer of the antibodies. The horseradish peroxidase-conjugated secondary antibodies (EMD Millipore; 1/4000) were diluted in TBST. Detection of the immunoreactive components on the blots were performed by using the ECL (Santa Cruz, CA, USA) Western Blotting detection reagent depending on the sensitivity of the antibodies.

Tseng H.L., Yang S.C., Yang S.H, & Shieh K.R. (2015). Hepatic Circadian-Clock System Altered by Insulin Resistance, Diabetes and Insulin Sensitizer in Mice. PLoS ONE, 10(3), e0120380.

+ Open protocol

+ Expand

Green Synthesis of Gold Nanoparticles

Check if the same lab product or an alternative is used in the 5 most similar protocols

We procured chloroauric acid (HAuCl₄) and sodium citrate from HiMedia Laboratories Pvt., Ltd. (Maharashtra, India). We bought polyvinylpropylene from Sigma-Aldrich, USA. C. verum bark was procured from local market. We received ELISA kits of IL-1β, IL-6, INF-γ, VEGF, and RANTES from R & D Systems, Canada. CD36, FAS, FABP4, PPARγ, and ACC antibodies were procured from Thermo Fisher Scientific and Abcam.

Sharma V.K., P.r.a.t.e.e.k.s.h.a., Gupta S.C., Singh B.N., Rao C.V, & Barik S.K. (2022). Cinnamomum verum-derived bioactives-functionalized gold nanoparticles for prevention of obesity through gut microbiota reshaping. Materials Today Bio, 13, 100204.

+ Open protocol

+ Expand

Isolation and Characterization of HBMEC Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols

For total protein isolation of cultured HBMECs, cells were washed twice with PBS prior to adding a lysis buffer. A plasma membrane protein extraction kit (Abcam, Cambridge, MA, USA) was used for sample collection. Protein samples (plasma membrane proteins or nuclear extractions) were equally separated in a 4%–20% NuPage gel prior to being transferred to nitrocellulose membranes. Primary antibodies used for overnight membrane incubation at 4 °C are as follows: PPARγ (1:1000, Thermo Fisher, Rockford, IL, USA), ZO-1 (1:1000, Cell Signaling, Beverly, MA, USA), VE-cadherin (1:1000, Enzo Life Sciences, Farmingdale, NY, USA), Na-K-ATPase (1:1000, Abcam), and Histone H3 (1:1000, Cell Signaling). Membranes were then incubated with horseradish peroxidase-linked secondary antibodies (1:2000) for 1 h at room temperature after being washed in PBS containing 0.1% Tween 20 and developed by enhanced chemiluminescence (Pierce). The optical density of protein bands was quantified with ImageJ.

Jiang Y., Lin L., Liu N., Wang Q., Yuan J., Li Y., Chung K.K., Guo S., Yu Z, & Wang X. (2020). FGF21 Protects against Aggravated Blood-Brain Barrier Disruption after Ischemic Focal Stroke in Diabetic db/db Male Mice via Cerebrovascular PPARγ Activation. International Journal of Molecular Sciences, 21(3), 824.

+ Open protocol

+ Expand

Western Blot Analysis of Cell Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols

Protein lysates from cultured cells or mouse tissues were analyzed by standard Western blot protocols [14 (link)] using the following primary antibodies: SCD1 (cat no.2438; Cell Signaling, Danvers, MA, USA), CD36 (cat no. PA1-46480; Thermo Fisher Scientific, Rockford, IL, USA), PPARγ (cat no. PA1-27585; Thermo Fisher Scientific), glyceraldehyde-3-phosphate dehydrogenase (cat no.2118; Cell Signaling), and actin (cat no. sc-1616; Santa Cruz Biotechnology, Inc, Santa Cruz, CA). Quantification of the proteins was performed using ImageJ software.

Geng T., Xia L., Russo S., Kamara D, & Cowart L.A. (2015). Prosteatotic genes are associated with unsaturated fat suppression of saturated fat-induced hepatic steatosis in C57BL/6 mice. Nutrition research (New York, N.Y.), 35(9), 812-822.

+ Open protocol

+ Expand

Peritoneal Protein Expression Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

The peritoneal tissue proteins were extracted using T-PER tissue protein extraction reagent added combined protease and phosphatase inhibitors (Thermo Fisher Scientific, Waltham, MA, U.S.A.). The protein concentrations were measured by BCA protein assay kit (Thermo Fisher Scientific) and equal amount of proteins were subjected to SDS-PAGE, then transferred to nitrocellulose membranes (Bio–Rad, Hercules, CA, U.S.A). After blocked with 5% nonfat milk, the membranes were incubated with antibodies against AQP-1 (1:500) (Santa Cruz, Dallas, Texas, U.S.A.), ZO-1 (1:500), PPAR-γ (1:500), p-PPAR-γ (1:500) (Thermo Fisher Scientific), and GAPDH (1:1,000) (Cell Signaling Technology, Beverly, MA, U.S.A.) respectively, followed by incubation with the HRP–conjugated secondary antibodies. Chemiluminescence was detected using ECL kit (GE Healthcare Life Sciences, Pittsburgh, PA, U.S.A.).

Zhang Y., Feng J., Wang Q., Zhao S., Xu J, & Li H. (2018). PPAR-γ agonist rosiglitazone ameliorates peritoneal deterioration in peritoneal dialysis rats with LPS-induced peritonitis through up-regulation of AQP-1 and ZO-1. Bioscience Reports, 38(3), BSR20180009.

+ Open protocol

+ Expand

Cellular Fractionation and Western Blot Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cytosolic and nuclear fractions of cells were separated using the Cell Fraction Kit (Abcam, Cambridge, UK). Nuclear or cytosolic proteins were separated by SDS-PAGE, transferred to polyvinylidene difluoride membranes, and incubated with primary antibodies against UCP1 (Sigma-Aldrich, St. Louis, MO, USA, cat# U6382), PPAR-γ (Thermo Fisher, Waltham, MA, USA, cat# ma5-14889), GAPDH (Sigma-Aldrich, St. Louis, MO, USA, cat# G8795), Histone H3 (Cell Signaling Technology, Danvers, MA, USA, cat# 4499P) or Lamin A (Abcam, Cambridge, UK, cat# ab8980). The membrane was then washed and incubated with the corresponding horseradish peroxidase-conjugated secondary antibodies (Cell Signaling Technology, Danvers, MA, USA Technology). Signals were visualized using an ECL detection kit (Thermo Scientific, cat# 34096). The relative expression of proteins was quantified using the ImageJ software.

Gou W., Wei H., Swaby L., Green E, & Wang H. (2023). Deletion of Spinophilin Promotes White Adipocyte Browning. Pharmaceuticals, 16(1), 91.

+ Open protocol

+ Expand

Gene Expression Analysis of Adipose Tissue

Check if the same lab product or an alternative is used in the 5 most similar protocols

The aqueous phase was mixed with an equal volume of 70% Ethanol before being loaded on an RNeasy mini column for extraction (Qiagen, Crawley, UK) as previously described (Walhin et al. 2013). Assays from Applied Biosystems were used: ADIPONECTIN (Hs00605917_m1), LEPTIN (Hs00174877_m1), SREBP‐1c (Hs01088691_m1), PDK4(Hs00176875_m1), FAS (Hs00188012_m1), PPARγ (Hs01115513_m1), GLUT4 (Hs00168966_m1), IRS2 (Hs00275843_s1), LPL (Hs01012567_m1), HSL (Hs00193510_m1), VISFATIN (Hs00237184_m1), IRS1 (Hs00178563_m1), IL18 (Hs00155517_m1), IL6 (Hs00985639_m1), AMPK (Hs01562315_m1 & Hs00178903_m1 combined). Real‐time PCR was performed using a StepOne^™ (Applied Biosystems, Warrington, UK). PPIA (Peptidylpropyl isomerase A) was used as an endogenous control. The comparative Ct method was used to process the data where ΔCt = Ct Target gene – Ct Endogenous control. Data were normalized to an internal calibrator and baseline.

Walhin J., Dixon N.C., Betts J.A, & Thompson D. (2016). The impact of exercise intensity on whole body and adipose tissue metabolism during energy restriction in sedentary overweight men and postmenopausal women. Physiological Reports, 4(24), e13026.

+ Open protocol

+ Expand

Quantifying Bone Cell Gene Expression

Check if the same lab product or an alternative is used in the 5 most similar protocols

Total RNA for Osx1‐TdRFP bone marrow cells was extracted from freshly isolated cells using Quick‐RNA™ MicroPrep (Zymo Research, Irvine, CA, USA) according to the manufacturer's instructions. Bone marrow‐derived stromal cells were cultured in 6‐well plates, and total RNA was extracted using TRIzol reagent (Invitrogen). cDNA was obtained from 1 to 2 μg of total RNA extract using the High‐Capacity cDNA Archive Kit (Applied Biosystems, Foster City, CA, USA) according to the manufacturer's instructions. TaqMan quantitative real‐time PCR was performed using the following primers from Applied Biosystems: Runx2 (Mm00501584_m1); Col1A1 (Mm00801666_g1); Pparγ (Mm00440945_m1); p16 (Mm00494449_m1); p21 (Mm00432448_m1); Tnf‐a (Mm00443258_m1); Il‐1α (Mm99999060_ m1); Il‐6 (Mm00446190_m1); Mmp‐13 (Mm00439491_m1); and Rankl (Mm00441908_m1). Osterix and Ocn mRNA levels were determined using custom‐made TaqMan Assay‐by‐Design primer sets 5′ATCTGACTT TGCTCCCCTTAACC3′ and 5′GGGCCCTGG TTGCAAGA3′; 5′GCTGCGC TCTGTCTCTCTGA3′ and 5′TGCTTGGACATGAAGGCT TTG3′, respectively. All reactions were run in triplicate, and target gene expression was calculated by normalizing to the housekeeping gene ribosomal protein S2 (Mm00475528_m1) using the ∆Ct method (Livak & Schmittgen, 2001).

Kim H., Chang J., Shao L., Han L., Iyer S., Manolagas S.C., O'Brien C.A., Jilka R.L., Zhou D, & Almeida M. (2017). DNA damage and senescence in osteoprogenitors expressing Osx1 may cause their decrease with age. Aging Cell, 16(4), 693-703.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!