Male Sprague-Dawley (SD) rats were purchased from Sina-British Sippr/Bk Lab Animal Ltd. (Shanghai, China). Tetramethylpyrazine (TMP), Crocin, Ferulic acid and Chlorogenic acid, acetylcholine, phenylephrine (PE) and D-glucose were purchased from Sigma-Aldrich Co. (St Louis, Missouri, USA). Dulbecco's modified Eagle's medium (DMEM) supplemented with 5.6 mmol/L glucose was obtained from Hclone (Logan, UT, USA). DMEM without red phenol, fetal bovine serum (FBS) and Trizol were purchased from Invitrogen (Carlsbad, CA, USA). CM-H2DCFDA, DAF-FM diacetate and MitoSOX™ Red mitochondrial superoxide indicators were obtained from Invitrogen (Carlsbad, CA, USA). JC-1 indicator was bought from Molecular Probes (Eugene, OR, USA). Antibodies including anti-complex III, anti-PGC-1α, anti-SIRT1 and anti-β-actin were purchased from Proteintech (ProteinTech Group, Chicago, IL, USA). Primers for PGC-1α, NRF-1, TFAM, SIRT1 and 18s were provided by Sangon Biotech CO. Ltd. (Shanghai).
Sirt1
Manufactured by Sangon
Sourced in China
SIRT1 is a nicotinamide adenine dinucleotide (NAD+)-dependent protein deacetylase enzyme. It plays a role in the regulation of cellular processes such as gene expression, metabolism, and stress response.
Automatically generated - may contain errors
Lab products found in correlation
Pgc 1α, by Sangon (2 mentions)
β actin, by Sangon (2 mentions)
Pparα, by Sangon (2 mentions)
D glucose, by Merck Group (1 mentions)
Chlorogenic acid, by Merck Group (1 mentions)
Ferulic acid, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Trizol, by Thermo Fisher Scientific (1 mentions)
Cm h2dcfda, by Thermo Fisher Scientific (1 mentions)
Daf fm diacetate, by Thermo Fisher Scientific (1 mentions)
Acetylcholine, by Merck Group (1 mentions)
Crocin, by Merck Group (1 mentions)
Phenylephrine, by Merck Group (1 mentions)
Anti β actin, by Proteintech (1 mentions)
Anti sirt1, by Proteintech (1 mentions)
Tetramethylpyrazine, by Merck Group (1 mentions)
Anti pgc 1α, by Proteintech (1 mentions)
Transscript first strand cdna synthesis supermix, by Transgene (1 mentions)
Axyprep multisource rna miniprep kit, by Corning (1 mentions)
Tnf α, by Sangon (1 mentions)
4 protocols using sirt1
1
Tetramethylpyrazine Enhances Mitochondrial Function
2
Comprehensive Gene Expression Analysis
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Total RNA was extracted with an Axyprep multisource RNA miniprep kit (Axygen, America), and cDNA was synthesized using TransScript First-Strand cDNA Synthesis SuperMix (TransGen Biotech, Beijing). Quantitative real-time PCR was performed using a SYBR Green kit (TransGen Biotech, Beijing) with a StepOnePlus real-time PCR system (ABI), and the primer sets used for MTTP, ApoA1, ApoB, ApoC2, CD31, TGFβ, TSP1, VEGFR1, IL-1β, TNFα, IL-6, IL-10, CCL2, PPARα, SIRT1, and β-actin (Sangon Biotech, Shanghai) are listed in Table 1 . Relative gene expression was measured with triplicates for each sample and normalized to β-actin.
3
Western Blot Analysis of Protein Expression
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Total protein from MMCs was extracted by ice-cold RIPA lysis buffer (Beyotime, Shanghai, China). Equal amount of protein samples were subjected into 10% SDS-PAGE (Beyotime, Shanghai, China), transferred onto 0.22 µm PVDF membranes (Millipore, Temecula, CA), and blocked in 5% nonfat milk. Then the membranes were incubated with specific primary antibodies. The primary antibodies used in the study were the following: cyclin D1 (Abcam, USA, 1:10000), p21 (Boster, China, 1:400), Col-4 (Proteintech, USA, 1:1000), FN (Proteintech, USA, 1:1000), HIF-1α (Sangon,China, 1:500), Sirt1 (Sangon, China, 1:500), TGF-β1 (Proteintech, USA, 1:500), and GAPDH (Abcam, USA, 1:5000). ECL system (Millipore, Temecula, CA) was used to detect the immunoreactive bands and GAPDH antibody was used as control. Gray value of protein bands were quantified by Image Lab software.
4
Comprehensive Metabolic Protein Analysis
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The BCA protein content detection kit was bought from KeyGen Biotech (Nanjing, China). Hypersensitive ECL chemiluminescence kit, protease, and phosphatase inhibitor were bought from NCM Biotech (Suzhou, China). The protein extraction kit was obtained from Beyotime Institute of Biotechnology (Shanghai, China). Non-esterified fatty acids (NEFA) and blood lipid test kits were purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). Revert Aid First Strand cDNA Synthesis Kit was purchased from Thermo Fisher Scientific (Waltham, MA, United States). Power SYBR Green PCR master mix was from Invitrogen (Carlsbad, CA, United States). The primers of AMPKα, SIRT1, PGC-1α, PPARα, GLUT4, UCP3, and β-actin were designed and synthesized by Sangon Biotech (Shanghai, China). Both primary antibodies to AMPKα, PGC-1α, PPARα, GLUT4, UCP3, GAPDH, Lamin B1, and Na, K-ATPase (Cat no: 66536-1-Ig, 66369-1-Ig, 15540-1-AP, 66846-1-Ig, 10750-1-AP, 60004-1-Ig, 66095-1-Ig, and 14418-1-AP) and secondary antibodies (Cat no: SA00001-1 and SA00001-2) were purchased from the Proteintech Group (Chicago, IL, United States). The antibody to Phospho-AMPKα (Thr172, Cat #: 50081S) was purchased from the Cell Signaling Technology (Boston, MA, United States). The antibody to SIRT1 (Cat #: ab189494) was purchased from Abcam (Cambridge, United Kingdom).
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