Immunoblotting were performed as described previously [13 ]. The antibodies used were : SAMM50 (dilution 1 : 1000, ab133709; Abcam, Cambridge, UK), β‐tubulin (dilution 1 : 8000; 10094‐1‐AP; Protein Tech, Wuhan, China), CPT1A (dilution 1 : 1000; A5307, ABclonal, Wuhan, China), 3‐ketoacyl‐CoA thiolase (dilution 1 : 1000; A15778; ABclonal), enoyl‐CoA hydratase (dilution 1 : 1000; 66117‐1‐Ig; Protein Tech) and long‐chain specific acyl‐CoA dehydrogenase (dilution 1 : 1000; 17526‐1‐AP; Protein Tech).
Cpt1a
Manufactured by ABclonal
Sourced in China
CPT1A is a laboratory protein that plays a key role in the transport of fatty acids into the mitochondria for energy production. It is an essential component in the study of lipid metabolism and related cellular processes.
Automatically generated - may contain errors
Lab products found in correlation
β tubulin, by Proteintech (1 mentions)
Uqcrc1, by Thermo Fisher Scientific (1 mentions)
Atp synthase subunit alpha, by Thermo Fisher Scientific (1 mentions)
Srebp1, by Santa Cruz Biotechnology (1 mentions)
Pparα, by Santa Cruz Biotechnology (1 mentions)
Cox 4, by Thermo Fisher Scientific (1 mentions)
Reverse transcription system kit, by Promega (1 mentions)
Sybr green, by Takara Bio (1 mentions)
Pgc 1α, by OriGene (1 mentions)
Pparα, by Bioworld Technology (1 mentions)
P ampk, by Bioworld Technology (1 mentions)
Acsl1, by ABclonal (1 mentions)
P acc, by Cell Signaling Technology (1 mentions)
β actin, by Santa Cruz Biotechnology (1 mentions)
P ampk, by ABclonal (1 mentions)
Cyp7a1, by ABclonal (1 mentions)
Hematoxylin and eosin h e, by Wuhan Servicebio Technology (1 mentions)
Gapdh, by ABclonal (1 mentions)
Horseradish peroxidase hrp conjugated goat anti rabbit igg, by ABclonal (1 mentions)
Superkine west femto maximum sensitivity substrate, by Abbkine (1 mentions)
6 protocols using cpt1a
1
Immunoblotting of Mitochondrial Proteins
2
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).
3
Quantitative and Western Blot Analysis of Lipid Metabolism Regulators
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Quantitative PCR (qPCR) was carried out as previously described [53 (link)]. All the primers used in these analyses were synthetized in Sangon Biotechnology Ltd. (Shanghai, China). Their sequences are shown in Table 1 .
Western blot analysis was performed as previously described [53 (link)]. β-Actin was employed as an internal reference. The primer antibodies for HTGL, MGL, HSL, p-HSL, AMPK, p-AMPK and PPARα were purchased from Bioworld (Shanghai, China). The primary antibodies for ACSL1, AOX and CPT1A were all purchased from ABclonal (Wuhan, China). Primary antibodies for β-actin and MSS4, as well as HRP-conjugated goat anti-rabbit and rabbit anti-mouse IgG, were purchased from Santa Cruz (Dallas, TX, USA).
Western blot analysis was performed as previously described [53 (link)]. β-Actin was employed as an internal reference. The primer antibodies for HTGL, MGL, HSL, p-HSL, AMPK, p-AMPK and PPARα were purchased from Bioworld (Shanghai, China). The primary antibodies for ACSL1, AOX and CPT1A were all purchased from ABclonal (Wuhan, China). Primary antibodies for β-actin and MSS4, as well as HRP-conjugated goat anti-rabbit and rabbit anti-mouse IgG, were purchased from Santa Cruz (Dallas, TX, USA).
4
Protein Expression Analysis in MDA-MB-231 Cells
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MDA-MB-231 cells were cultured or administrated for 36 h and then lyzed using lysis buffer and the total protein samples were isolated and eluted with SDS buffer, separated by SDS-polyacrylamide gels, and electroblotted onto PVDF membranes. The specific protein bands were detected with Odyssey Scanning System (LI-COE Inc., Superior St., Lincoln, NE) according to the previous report [27] (link). The following are the origins of the primary antibodies and the corresponding diluting proportions. (1) G6PD from Abclonal Technology, 1:2000, was used to determine the level of PPP; (2) β-actin from Santa Cruz Biotechnology, 1:2000, was used as internal reference; (3) p-ACC from Cell Signaling Technology, 1:1000, was used to determine the phosphorylation of ACC and the activation of AMPK; (4) CPT1A from Abclonal Technology, 1:2000, was used to determine the level of FAO; (5) PDH from Cell Signaling Technology, 1:1000, was used to determine the level of acetyl-CoA produced from glucose; (6) LDH from Abclonal Technology, 1:2000, was used to determine the level of glycolysis; (7) AMPK from Abclonal Technology, 1:2000, was used to determine the expression level of AMPK and (8) p-AMPK from Abclonal Technology, 1:2000, was used to determine the level of AMPK activation. Western blot assays for each protein were performed at least three times
5
Torularhodin Isolation and Purification from Sporidiobolus pararoseus
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Torularhodin (purity > 95%) was isolated and purified from the extract of Sporidiobolus pararoseus (JD-2 CCTCC M 2010326) according to a previously published method [10 (link)]. Animal diets were purchased from TROPHIC Animal Feed High-tech Co., Ltd. Commercial kits for serum concentration of total cholesterol (TC), low-density lipoprotein cholesterol (LDL-c), high-density lipoprotein cholesterol (HDL-Cc), and triglyceride (TG) were purchased from Jiancheng Technology Co. (Nanjing, China). Hematoxylin and eosin (H&E) were obtained from Servicebio Technology Co. (Wuhan, China). Antibodies of PPARα, PPAR-γ, CYP7A1, CPT1A, SLC27A4, and GAPDH were obtained from ABclonal Technology Co. Ltd. (Wuhan, China). Western fluorescence detection reagent was obtained from Beyotime Biotechnology (Shanghai, China).
6
Molecular Mechanisms of Cisplatin-Induced Apoptosis
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Cisplatin (#T1564) is from Topsicence (United States); FPS-ZM1 (#HY-19370) is from MedChemExpress (United States); rabbit anti-RAGE (#ab3611) is from Abcam (United Kingdom); rabbit anti-Bax (#CPA1092), and Bcl-2 (#CPA1095) antibodies are from Cohesion Bioscience (United Kingdom); rabbit anti-Phospho-NF-κB p65 (Ser536) (#3033) and NF-κB p65 (#3034) antibodies are from Cell Signaling Technology (United States); rat anti-F4/80 antibody (#14-4801-82) is from Invitrogen United States; mouse anti-GAPDH (#AC033), rabbit anti-β-actin (#AC026), Cpt1a (#A5307) and PGC-1α (#A12348) and horseradish peroxidase (HRP)-conjugated goat anti-rabbit IgG (#AS014) antibodies are from ABclonal (China). One Step Terminal transferase dUTP nick-end labelling (TUNEL) Apoptosis Assay Kit (#C1090) is from Beyotime (China). SuperKine™ West Femto Maximum Sensitivity Substrate (#BMU102-CN) is from Abbkine (China).
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