Pgc 1a

Manufactured by Abcam

Sourced in United States

PGC-1a is a transcriptional coactivator that regulates genes involved in energy metabolism. It is a key regulator of mitochondrial biogenesis and respiration.

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Lab products found in correlation

β actin, by Abcam (2 mentions) Sirt1, by Abcam (1 mentions) P acc, by Abcam (1 mentions) Gadph, by Abcam (1 mentions) Nf kb, by Abcam (1 mentions) P ampk, by Abcam (1 mentions) Rhea ecl, by US Everbright (1 mentions) Ripa lysate, by Keygen Biotech (1 mentions) Fabp4, by Abcam (1 mentions) Cd140a, by R&D Systems (1 mentions) Rab27a, by R&D Systems (1 mentions) Srx 101a, by Konica Minolta (1 mentions) C ebpα, by Santa Cruz Biotechnology (1 mentions) Hrp conjugated secondary antibody, by Cell Signaling Technology (1 mentions) Plin1, by Santa Cruz Biotechnology (1 mentions) Tgx gel, by Bio-Rad (1 mentions) Pparg, by Santa Cruz Biotechnology (1 mentions) Trans blot turbo, by Bio-Rad (1 mentions)

4 protocols using pgc 1a

Western Blot Analysis of Metabolic Regulators

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Tissues were homogenized and lysed by RIPA buffer. After centrifugation of the lysates at 12,000 g for 10 min at 4 degrees, the supernatants were collected. Equal amounts of protein were electrophoresed on SDS-polyacrylamide gel and transferred to a nitrocellulose membrane. The membrane was then preincubated with blocking solution (PBST containing 5% fat-free milk) for 2h before incubation with primary antibodies at 4 degrees overnight. The following primary antibodies were used: GADPH (1:4000; Abcam), β-actin (1:4000; Abcam), AMPK, (1:2000; Abcam). p-AMPK (1:2000; Abcam), ACC (1:2000; Abcam), p-ACC (1:2000; Abcam), SIRT1(1:2000; Abcam), PGC1a (1:2000; Abcam), NF-kB (1:2000; Abcam). After washing with PBST, the blot was incubated with secondary antibody (HRP-conjugated, 1:10,000 dilution) for 90 min and then washed and detected by an enhanced chemiluminescence detection kit (Haigene).

Diao Y., Nie J., Tan P., Zhao Y., Zhao T., Tu J., Ji H., Cao Y., Wu Z., Liang H., Huang H., Li Y., Gao X, & Zhou L. (2020). Long-term low-dose ethanol intake improves healthspan and resists high-fat diet-induced obesity in mice. Aging (Albany NY), 12(13), 13128-13146.

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Mitochondrial Protein Expression Analysis

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Total protein was extracted with RIPA lysate (KeyGEN BioTECH, Nanjing, China) and phenylmethylsulfonyl fluoride (PMSF; KeyGEN BioTECH). Samples (60 μg, 5–10 μL) were run on an SDS-PAGE gel followed by blotting to a polyvinylidene fluoride (PVDF) membrane. After blocking with 5% skim milk, the membranes were incubated with the following primary antibodies: β-actin (1:5000 Abcam, Branford, CT, USA), PGC-1a (1:1000; Abcam), NRF-1 (1:1000; Abcam), and Tfam (1:2000; Abcam), fusion protein mitofusin 1 (Mfn-1) (1:2000; Abcam), optic atrophy 1 (OPA-1) (1:1000; Abcam) and dynamin-related protein 1 (DRP-1) (1:500; Abcam), followed by incubation with appropriate secondary antibodies. Rhea ECL (US Everbright Inc., Suzhou, China) was used as the developer reagent, and the band intensity was assessed by using Image lab software and referenced to β-actin.

Shao Q., Meng L., Lee S., Tse G., Gong M., Zhang Z., Zhao J., Zhao Y., Li G, & Liu T. (2019). Empagliflozin, a sodium glucose co-transporter-2 inhibitor, alleviates atrial remodeling and improves mitochondrial function in high-fat diet/streptozotocin-induced diabetic rats. Cardiovascular Diabetology, 18, 165.

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Immunofluorescent Imaging of Cellular Organelles

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Cells were fixed on coverslips with 2% paraformaldehyde for 15 minutes and then rinsed with cold PBS. Liver tissue harvested from mice was removed after whole body perfusion with cold PBS followed by 2% paraformaldehyde. Tissue was placed in 2% paraformaldehyde for 1 hour and then switched to 30% sucrose in distilled water for 12 hours. Tissue was then slowly frozen in 2-methylbutane. Tissue sections were obtained at 7 μm and were then stained using 2′-(4-hydroxyphenyl)-5-(4-methyl-1-piperazinyl)-2,5′-bi(1H-benzimidazole) trihydrochloride (Sigma, cat no. B-2883) for nuclear identification and 488-conjugated phalloidin antibody (BioLegend, cat no. 424201) for actin cytoskeletal identification. Specific antibodies to LC3 (Novus, St. Charles, MO, USA), PGC-1a (Abcam, Cambridge, MA, USA), NRF1, NRF2, or HO-1 (Abcam, Cambridge, MA, USA) were additionally utilized for imaging as indicated according to manufacturer's instructions. All slides were scanned under the same conditions for magnification, exposure time, lamp intensity, and camera gain. Confocal images were acquired using the Nikon A1 with a PlanApo N (×20 with and without a 2-fold digital zoom). All imaging studies were repeated at least n = 3 times on biological replicates.

Cyr A., Chambers L., Waltz P.K., Whelan S.P., Kohut L., Carchman E., Dyer M., Luciano J., Kautza B., Gomez H.D., Otterbein L.E., Rosengart M.R., Shiva S, & Zuckerbraun B.S. (2019). Endotoxin Engages Mitochondrial Quality Control via an iNOS-Reactive Oxygen Species Signaling Pathway in Hepatocytes. Oxidative Medicine and Cellular Longevity, 2019, 4745067.

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Immunoblotting for Adipogenic Markers

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For immunoblotting, 25–50μg of protein was loaded onto a 10% TGX gel (Bio-Rad) and separated at 200 volts for ~30 to 40 minutes. Protein was transferred to a PVDF membrane using Bio-Rad Trans-blot Turbo (1.5amps, 15 volts for 30 minutes). Membranes were blocked using 5% w/v non-milk dissolved in PBS with 0.1% tween-20 (PBS-T) for 30 minutes and incubated with primary antibody overnight at 4°C. Primary antibodies and dilutions used were as follows: from Cell Signaling Technologies - ACTB 1:5000 (#4970S), C/EBPα 1:750 (#8178S), FASN 1:1000 (#3180S), PLIN1 1:4000 (#9349S), PPARG 1:500 (#2443S), TUBA1B 1:2500 (#2125S); from Santa Cruz Biotechnology - CD73 1:1000 (#sc-398260), CD140A 1:1000 (#sc-398206), Rab5 1:750 (#sc130010), Rab27a 1:1000 (#sc136996); from R&D Systems - UCP1 1:750 (#MAB6158-SP); and from Abcam - FABP4 1:2500 (#ab92501), PGC1A 1:2000 (#ab54481). The next day, the membranes were washed 3× for 15 minutes each in PBS-T and incubated in HRP-conjugated secondary antibody from Cell Signaling Technology (#7074S and #7076S) diluted 1:4000 in PBT-T supplemented with 5% w/v milk for 1h. Membranes were washed 3× for 15 minutes each and developed using Immobilon forte chemiluminescent reagent (Millipore cat #WBLUF0100). Signal was detected using HyBlotCL ® autoradiography films (Denville #E3212–1001371) in a dark room and developed on a Konica developer (SRX-101A).

Boucher J.M., Robich M., Scott S.S., Yang X., Ryzhova L., Turner J.E., Pinz I, & Liaw L. (2018). Rab27a regulates human perivascular adipose progenitor cell differentiation. Cardiovascular drugs and therapy, 32(5), 519-530.

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