Pgc 1β

Manufactured by Santa Cruz Biotechnology

Sourced in United States

PGC-1β is a protein that plays a role in the regulation of mitochondrial biogenesis and metabolism. It is a transcriptional coactivator that interacts with various transcription factors to promote the expression of genes involved in energy production and oxidative metabolism. PGC-1β is expressed in tissues with high energy demands, such as skeletal muscle, heart, and liver.

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Mouse anti-Pgc-1β (1 citations)

3 protocols using pgc 1β

Mitochondrial Energy Metabolism Gene Expression

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Following total RNA extraction using TRIzol (Invitrogen, NY), cDNA was synthesized from total RNA by using RT² First Strand kit and protocol (Qiagen, CA). Real time PCR was conducted using Bio-Rad iCycler (iCycler iQ Multi-Color Real Time PCR Detection System; Bio-Rad, CA) real time system. The expression of human and rat mitochondrial energy metabolism genes involved in mitochondrial respiration was profiled by using Mitochondrial Energy Metabolism plus RT² Profiler PCR Array (Qiagen). Data analysis was performed by using the Web-based software for cataloged and custom arrays developed by Qiagen (http://pcrdataanalysis.sabiosciences.com/pcr/arrayanalysis.php). Protein extraction and western blot analysis using U87 and B12 cells were performed as described earlier (Baarine et al. 2009 (link)) using various antibodies raised against PGC-1α (Abcam 106814, MA), PGC-1β (Santa-Cruz, TX), ABCD1 (EMD Millipore, MA), β-actin (Cell Signaling, MA), and GAPDH (Abcam).

Baarine M., Beeson C., Singh A, & Singh I. (2015). ABCD1 deletion-induced mitochondrial dysfunction is corrected by SAHA: implication for adrenoleukodystrophy. Journal of neurochemistry, 133(3), 380-396.

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Liver Protein Quantification via Western Blot

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Liver proteins were quantified by western blot analysis as follows using BioRad, (Hercules, CA) Criterion and ChemiDoc systems following manufacturers’ instructions. PGC1α antibody was purchased from EMD Millipore, Billerica, MA, Cat# ST1202. All other antibodies purchased from Santa Cruz Biotechnology, Santa Cruz, CA were as follows: FOXO1Ser256 Cat# sc-101681; G6Pase-α Cat# sc-27198; PEPCK Cat# sc-32879; PPARα Cat# sc-9000; SREBP-1 Cat# sc-13551; mTORC Cat# sc-8319; PPARγ Cat# sc-7273; PGC1β Cat# sc-67286, NFκB p65 Cat# sc-8008; IKKαβ Cat# sc-7607; SOCS3 Cat# sc-9023; JNK Cat# sc-571; Actin Cat# sc-47778. Criterion gradient tris-glycine precast gels, secondary antibodies, PVDF blotting membranes, molecular weight markers, and Enhanced Chemiluminescence (ECL) reagents were also purchased from BioRad. Samples from 8 SHR Vehicle, 8 SHR Bromocriptine treated rats, and 6 Wistar wild type controls were loaded onto the same 26 well Criterion gel along with the molecular weight markers; band intensity was compared only within the samples loaded onto the same gel. A housekeeping protein (Actin) was concurrently quantified on all gels, and the test protein amount was normalized to Actin in the Western blot analysis. Protein bands were quantified with BioRad ImageLab 4.1 software.

Ezrokhi M., Luo S., Trubitsyna Y, & Cincotta A.H. (2014). Neuroendocrine and metabolic components of dopamine agonist amelioration of metabolic syndrome in SHR rats. Diabetology & Metabolic Syndrome, 6, 104.

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Immunoblotting Analysis of Muscle-Related Proteins

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Tissue extracts were prepared in radioimmunoprecipitation assay (RIPA) buffer (Cell Signaling Technology, USA). The resulting preparations (10 to 20 μg) were then fractionated by polyacrylamide-SDS gel electrophoresis and immunoblotted with SIRT-1 (sc-15404), PGC-1β (sc-373771), ATP5B (sc-55597), MuRF1 (sc-32920), MAFbx (sc-33782), tropomyosin (sc-28543), MYH (sc-20641), hypoxanthine phosphoribosyltransferase (HPRT) (sc-20975), NOX2 (gp91-phox) (sc-130543), and β-actin (47778) from Santa Cruz Biotechnology, Inc., USA; with UCP-3 (catalog no. 97000), pAMPKβ1 (Ser182) (catalog no. 4186), and AMPKβ1 (catalog no. 4150) from Cell Signaling Technology, USA; with ATG5 (NBP2-24389) and LC3B (NB100-2220) from Novus Biologicals, USA; and with anti-SQSTM1/p62 (catalog no. ab56416) from Abcam, USA. The primary and secondary antibodies were used at dilutions of 1:1,000 and 1:10,000, respectively. The blots were visualized with SuperSignal West Pico chemiluminescent substrate (Thermo Scientific, Rockford, IL, USA), according to the manufacturer’s instructions. The immunoreactive bands were analyzed using ImageJ software, and a densitometry analysis was conducted.

Bandyopadhaya A., Tzika A.A, & Rahme L.G. (2019). Pseudomonas aeruginosa Quorum Sensing Molecule Alters Skeletal Muscle Protein Homeostasis by Perturbing the Antioxidant Defense System. mBio, 10(5), e02211-19.

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