The antibodies used for western blots in this study are as follows. BMAL1 (ab93806, Abcam, 1:10000), Acetyl-BMAL1 (AB15396, EMD Millipore, 1:1000), REV-ERBα (#13418, Cell Signaling Technology, 1:1000), PER2 (PER21-A, Alpha Diagnostic International, 1:1000), CRY1 (A302–614A, Bethyl Laboratories, 1:2000), GR (#12041, Cell Signaling Technology, 1:2000), Phospho-CREB (#9198, Cell Signaling Technology, 1:1000), CREB (#9197, Cell Signaling Technology, 1:1000), FOXO1 (#2880, Cell Signaling Technology, 1:1000), PPARα (sc-9000, Santa Cruz Biotechnology, 1:1000), SIRT1 (07–131, EMD Millipore, 1:10000), PPARδ (PA1–823A, Thermo Fisher Scientific, 1:1000), TFEB (A303–673A, Bethyl Laboratories, 1:2000), ACTIN (ab3280, Abcam, 1:10000), a—TUBULIN (T5168, SIGMA-ALDRICH, 1:10000), p84 (GTX70220, GeneTex, 1:10000), secondary antibodies (12–348 and AP160P, EMD Millipore, 1:10000). The antibodies used for ChIP are as follows. BMAL1 (ab93806, Abcam, 2 µg), GR (#12041, Cell Signaling Technology, 5 µl), CREB (sc-186 X, Santa Cruz Biotechnology, 10 µg), PPARα (sc-9000 X, Santa Cruz Biotechnology, 8 µg), normal rabbit IgG (sc-2027, Santa Cruz Biotechnology).
Pparα
Manufactured by Santa Cruz Biotechnology
Sourced in United States
PPARα is a nuclear receptor that regulates the expression of genes involved in lipid metabolism and energy homeostasis. It plays a key role in the control of fatty acid oxidation and ketogenesis.
Automatically generated - may contain errors
Lab products found in correlation
Pparγ, by Santa Cruz Biotechnology (15 mentions)
β actin, by Santa Cruz Biotechnology (12 mentions)
Srebp1, by Santa Cruz Biotechnology (6 mentions)
β actin, by Merck Group (5 mentions)
Srebp 1c, by Santa Cruz Biotechnology (4 mentions)
β actin, by Abcam (4 mentions)
P ampkα, by Cell Signaling Technology (3 mentions)
P acc, by Cell Signaling Technology (3 mentions)
Ampkα, by Cell Signaling Technology (3 mentions)
Pgc 1α, by Santa Cruz Biotechnology (3 mentions)
Pvdf membrane, by Bio-Rad (3 mentions)
Quantity one software, by Bio-Rad (3 mentions)
Pparα, by Merck Group (3 mentions)
Alexa fluor 594 conjugated goat anti rabbit igm, by Thermo Fisher Scientific (3 mentions)
Duolink proximity ligation assay kit, by Merck Group (3 mentions)
Alexafluor 488 conjugated goat anti mouse igg1, by Thermo Fisher Scientific (3 mentions)
Ncor1, by Cell Signaling Technology (3 mentions)
Gapdh, by Santa Cruz Biotechnology (3 mentions)
P mtor, by Cell Signaling Technology (2 mentions)
β actin, by Cell Signaling Technology (2 mentions)
67 protocols using pparα
1
Antibodies for Circadian Clock Proteins
2
Western Blot Analysis of Tissue Proteins
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Western blot analysis was performed with the same method used in a previous study [25 (link)]. Cytosolic and nuclear proteins were extracted from the liver, gastrocnemius tissue, and hippocampus with lysis buffers containing protease/phosphatase inhibitor, cytosol-extracting buffer. A total of 30 μg of protein for each group was used for western blot analysis. The extract was separated by 8~12% SDS-PAGE gel electrophoresis then transferred to a polyvinylidene fluoride membrane (Millipore, Billerica, MA, USA). Then, it was incubated overnight with primary antibodies and probed with the respective secondary antibody reagent (Biorad, Hercules, CA, USA). The bands were adjusted by the band levels of α-tubulin (cytosol) or PCNA (nucleus) [25 (link)].
Antibody list: pAkt (sc-81434), Akt (sc-514032), GLUT4 (sc-53566), GLUT2 (sc-518022), FGF21 (sc-81946), KYN (sc-69890), PPARα (sc-398394), FGFR1 (sc-57132), CTSB (sc-365558), PGC1α (sc-517380), Foxo3a (sc-48348), Atroin-1 (sc166806), Murf-1 (sc-398608) (Santa Cruz Biotechnology, CA, USA, 1:200), IRS-1 (#2382), pIRS-1 (#2381), mTOR (#2972), pmTOR (#2971), LC3 (#2775), (#2535), AMPK (#2532) (cell signaling technology, MA, USA, 1:2000), FNDC5 (ab131390), BDNF (ab108319), β-klotho (ab127879) (Abcam, Cambridge, MA, USA, 1:10,000), α-tubulin (T5168, Signa Aldrich, MO, USA, 1:4000), PCNA (610665, Enzolife science, Farmingdale, NY, USA, 1:1000).
Antibody list: pAkt (sc-81434), Akt (sc-514032), GLUT4 (sc-53566), GLUT2 (sc-518022), FGF21 (sc-81946), KYN (sc-69890), PPARα (sc-398394), FGFR1 (sc-57132), CTSB (sc-365558), PGC1α (sc-517380), Foxo3a (sc-48348), Atroin-1 (sc166806), Murf-1 (sc-398608) (Santa Cruz Biotechnology, CA, USA, 1:200), IRS-1 (#2382), pIRS-1 (#2381), mTOR (#2972), pmTOR (#2971), LC3 (#2775), (#2535), AMPK (#2532) (cell signaling technology, MA, USA, 1:2000), FNDC5 (ab131390), BDNF (ab108319), β-klotho (ab127879) (Abcam, Cambridge, MA, USA, 1:10,000), α-tubulin (T5168, Signa Aldrich, MO, USA, 1:4000), PCNA (610665, Enzolife science, Farmingdale, NY, USA, 1:1000).
3
Protein Expression Analysis of Troglitazone-Treated Cells
4
Immunoblotting Techniques for Lipid Metabolism
5
Hepatic Metabolic Regulation Protocol
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Chemicals were purchased from Sigma–Aldrich (St. Louis, MO, USA) unless indicated otherwise. Antibodies against phosphorylated JNK, total JNK, phosphorylated IκBα, total IκBα, PPARα, and CREBH were purchased from Santa Cruz Biotechnologies, Inc. (Santa Cruz, CA, USA). Antibodies against GRP78, IRE1a, Xbp1s, β-actin and the secondary antibodies were purchased from Cell Signaling Technologies (Danvers, MA, USA). Antibodies against FGF21 were purchased from Abcam (Boston, MA, USA). The kit for determining ALT, AST, and FFA were purchased from Abcam. The kit for determining TG, TC, and HDL weaspurchased from Fujifilm Wako Diagnostics USA (Mountain View, CA, USA). The glycogen enzyme-linked immunosorbent assay (ELISA) kit was purchased from BioAssay Systems (Hayward, CA, USA). The rabbit insulin ELISA kit was purchased from Crystal Chem (Elk Grove Village, IL, USA). The periodic acids staining kit, Gomori's trichrome staining kit were purchased from Fisher Scientific (Hampton, NH, USA). The assay kits and antibodies information are listed in Table S2 .
6
Liver Tissue Protein Analysis
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The protein concentration was determined in the supernatant of liver tissue. The samples with equal amounts of protein were analyzed for Phospho-Tyr705-STAT3 (Cell signaling Technology, Boston, MA), Phospho-217-C/EBPβ (Santa Cruz Biotechnologies, CA, USA), PGC-1α (Abcam plc, Cambridge, UK), Phospho-Ser32-IқBα (Cell signaling Technology, Boston, MA), PPARα (Santa Cruz Biotechnologies, CA, USA), ABCG5 (Santa Cruz Biotechnologies, CA, USA), ABCG8 (Novus Biological, Littleton, USA), CYP7A1 (Abcam plc, Cambridge, UK), CYP19A1 (Abcam plc, Cambridge, UK), β-actin (Abgent, San Diego, CA, USA) and GAPDH (CWBIO, Beijing, China) by SDS-PAGE and Western blotting.
7
Immunoblot Analysis of Cellular Proteins
8
Metabolic Regulation in Murine Obesity
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Antibodies list: CPT1A (A5307, ABclonal, Wuhan, China), UCP1 (A5857, ABclonal), UCP3(A16996, ABclonal), PPARα (sc-9000, Santa Cruz, Dallas, TX, USA), PPARγ (2435S, CST, Danvers, MA, USA), FAS (3189S, CST), SREBP1 (Sc13551, Santa Cruz), PGC-1α (TA319007, Origene, Rockville, MD, USA), NDUFS3 (459130, Invitrogen, Waltham, MA, USA), SDHB (459230, Invitrogen), UQCRC1 (459140, Invitrogen), COX4 (459600, Invitrogen), ATP Synthase Subunit Alpha (459240, Invitrogen), SOD2 (sc-137254, Santa Cruz), β-Actin (3700S, CST), α-Tubulin (3873S, CST).
Mice diet was provided by SLAC Laboratory Animal Co. Ltd. (Shanghai, China). Insulin was purchased from Nove Nordisk A/S. The blood glucose meter and blood glucose test strip were both purchased from ROCHE (ACCU-CHEK Active).
The Reverse Transcription System kit was purchased from Promega; SYBR green was purchased from Takara; polymerase chain reaction (PCR) primers were synthesized by Beijing Qingke biotechnology Co. Ltd. Nitrocellulose membranes used in W.B. were purchased from PerkinElmer Life Sciences. Other reagents used in this study were purchased from Sigma (St. Louis, MO, USA).
Mice diet was provided by SLAC Laboratory Animal Co. Ltd. (Shanghai, China). Insulin was purchased from Nove Nordisk A/S. The blood glucose meter and blood glucose test strip were both purchased from ROCHE (ACCU-CHEK Active).
The Reverse Transcription System kit was purchased from Promega; SYBR green was purchased from Takara; polymerase chain reaction (PCR) primers were synthesized by Beijing Qingke biotechnology Co. Ltd. Nitrocellulose membranes used in W.B. were purchased from PerkinElmer Life Sciences. Other reagents used in this study were purchased from Sigma (St. Louis, MO, USA).
9
Liver Protein Expression Analysis
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According to the methods of our previous study, total proteins were isolated from 50 mg of liver tissue and western blotting was performed. The blots were probed using the following antibodies: HIF-1α (sc-10790, 1:1000), PPARα (sc-9000, 1:1000), SREBP-1 (2A4) (sc-13551, 1:500), DEC1 (s-8) (sc-101023, 1:1000), AMPKα1/2 (sc-74461, 1:1000), Thr172-p-AMPKα1/2 (sc-33524, 1:1000), and β-actin (sc-477778, 1:1000); the above-mentioned antibodies were all from Santa Cruz Biotechnology, Dallas, TX, USA. ACC (#3662, Cell Signaling Technology, Inc., Danvers, MA, USA) and Ser79-p-ACC (#3661; Cell Signaling Technology, Inc., Danvers, MA, USA). The density of protein bands was analyzed using Bio-Rad imaging software (Bio-Rad Laboratories, Hercules, CA, USA). The individual values were originally expressed as a ratio of a standard (β-actin content) and then expressed as a fold change of the control group value.
10
Protein Isolation and Western Blot Analysis
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Protein isolation and western blot analysis were performed as described in literature (Wu et al., 2010 (link)). Briefly, protein samples were placed in sodium dodecyl sulfate-polyacrylamide gel electrophoresis and blotted onto polyvinylidene difluoride membranes. The membranes were blocking with non-fat dry milk for 1 h and incubated at room temperature for 2 h with p-LKB1, LKB1, p-AMPKα, AMPKα, p-AMPKβ, AMPKβ, p-ACC, ACC (Cell Signaling Technology, Boston, MA, USA) or SREBP-1, CYP2E1, GAPDH, Sirt1, PPARγ (Abcam, Cambridge, MA, USA) or caspase-1, β-actin, PPARα (Santa Cruz, CA, USA) or IL-1β (R&D Systems Europe Ltd., Abingdon, UK). Then the membranes were followed by incubated with HRP-conjugated secondary antibody for 1 h at room temperature and visualized by ECL Prime Western Blotting Detection Reagent (Bio-Rad, USA).
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