Dulbecco's modified Eagle's medium (DMEM) and other culture reagents were obtained from Gibco Life Technologies (Grand Island, NY).The cell culture plates were purchased from Nalge Nunc International (Roskilde, Denmark). Human insulin (HumulinR) was from Eli Lilly S.A.S.(Fegersheim, France). Bovine serum albumin (BSA), forskolin, IBMX, and dexamethasone were purchased from Sigma (St Louis, MO, USA). Compound C was purchased from Calbiochem (San Diego, CA). Anti-CREB, anti-phospho-CREB (Ser133), anti-PPARγ, anti-C/EBPα, anti-fatty acid synthase (FAS), anti-fatty acid binding protein 4 (FABP4), anti-C/EBPβ, anti-AMPK, anti-phospho-AMPK (Thr172), anti-acetyl-CoA carboxylase (ACC), anti-phospho-ACC(Ser79), anti-β-actin, anti-α1-tubulin, anti-mouse IgG and anti-rabbit IgG conjugated with horseradish peroxidase were from Cell Signaling Technology (Beverly, MA, USA). Murine-derived 3T3-L1 preadipocytes were purchased from American Type Culture Collection (Rockville, MD). Berberine was obtained from the National Institute for the Control of Pharmaceutical and Biological Products (Beijing, China).
Anti fatty acid synthase
Manufactured by Cell Signaling Technology
Sourced in United States
Anti-fatty acid synthase (FAS) is a laboratory product that detects the presence of the fatty acid synthase enzyme. Fatty acid synthase is a multi-enzyme complex responsible for the synthesis of long-chain fatty acids. This product can be used to identify and quantify the levels of fatty acid synthase in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Anti acetyl coa carboxylase, by Cell Signaling Technology (4 mentions)
Anti erk, by Cell Signaling Technology (2 mentions)
Horseradish peroxidase, by Cell Signaling Technology (1 mentions)
Cell culture plates, by Thermo Fisher Scientific (1 mentions)
Human insulin humulinr, by Eli Lilly (1 mentions)
Anti phospho creb ser133, by Cell Signaling Technology (1 mentions)
Anti phospho ampk thr172, by Cell Signaling Technology (1 mentions)
Anti phospho acc ser79, by Cell Signaling Technology (1 mentions)
Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (1 mentions)
Anti pparγ, by Cell Signaling Technology (1 mentions)
3 isobutyl 1 methyl xanthine (ibmx), by Merck Group (1 mentions)
Anti ampk, by Cell Signaling Technology (1 mentions)
Anti creb, by Cell Signaling Technology (1 mentions)
Anti c ebpα, by Cell Signaling Technology (1 mentions)
Anti β actin, by Cell Signaling Technology (1 mentions)
Dexamethasone (dex), by Merck Group (1 mentions)
Anti hsl, by Cell Signaling Technology (1 mentions)
Anti p hsl, by Cell Signaling Technology (1 mentions)
Anti lc3 antibody nb100 2220, by Novus Biologicals (1 mentions)
Anti beclin 1 nb110 87318, by Novus Biologicals (1 mentions)
6 protocols using anti fatty acid synthase
1
Berberine's Effects on Adipocyte Differentiation
2
Adipocyte Lipolysis and Autophagy Regulation
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Forskolin (F6886), NH4Cl (A9434) and leupeptin (L2884) were purchased from Sigma–Aldrich (St. Louis, MO, USA). H89 (#371962) and KT5823 (#420321) were from Calbiochem (San Diego, CA, USA). Anti-perilipin 1 (#9349), anti-HSL (#4107), anti-pHSL (Ser552)(#4139), anti-pHSL (Ser554)(#4137), anti-pHSL (Ser650)(#4126), anti-acetyl-CoA carboxylase (#3676), anti-adiponectin (#2789), anti-CCAAT/enhancer-binding protein α (C/EBPα) (#8178), anti-fatty acid binding protein 4 (FABP4) (#3544) and anti-fatty acid synthase (FAS) (#3180), anti-p62/SQSTM1 (#5114s) antibodies were from Cell Signaling Technology (Beverly, MA, USA). Anti-beclin-1 (NB110-87318), anti-Atg5 (NB110-53818), and anti-LC3 antibody (NB100-2220) was purchased from Novus Biologicals (Littleton, CO, USA). Anti-GAPDH (SC-25778) antibody was from Santa Cruz Biotechnology (Dallas, TX, USA).
3
Extraction and Analysis of Liver Proteins
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Total proteins were extracted from liver tissue, homogenizing the samples in modified RIPA buffer [50 mM Tris–HCl pH 7.5, 150 mM NaCl, 1% Triton, 0.25% sodium deoxycholate, 10 mM sodium pyrophosphate, 1 mM sodium orthovanadate, 1 mM sodium fluorure (Sigma-Aldrich), and protease inhibitor cocktail (Roche Diagnostics GmbH)], and then centrifuging the supernatant at 10,000 g for 10 min at 4°C, twice (18 (link)). 15 µg of total protein was separated through SDS-polyacrylamide gel electrophoresis (run on 7 and 12%), as previously described (22 (link), 23 (link)). Protein expression was measured using the following specific antibodies: anti-sterol regulatory element-binding transcription factor 1 (SREBP-1; 1:500); anti-extracellular regulated MAP kinase (ERK; 1:1,000) and anti-phospho-ERK (Thr202/Tyr204 p-ERK; 1:500) (Santa Cruz Biotechnology); anti-fatty acid synthase (FAS; 1:1,000) (Cell Signaling Technology); and anti-GAPDH (1:40,000) (Thermo Fisher Scientific Inc.). Western blot (WB) detection system ECL Plus (Perkin Elmer) was used to visualize the immune complexes (24 (link)).
4
Liver Protein Analysis by Western Blot
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Protein isolation from liver tissue and Western blot analysis were performed as previously described (Dou et al., 2018 (link)). The following antibodies were used: anti-phosphorylated-AMPKα (Thr172), anti-AMPKα, anti-PPARα, anti-CPT-1, anti-acetyl-CoA-carboxylase (ACC), anti-phosphorylated-ACC, anti-SREBP-1c, anti-fatty acid synthase (FAS), anti-DGAT-2, anti-P53, anti-Bcl2, anti-Bax, anti-TLR4, anti-JNK, anti-phosphorylated-JNK, anti-phosphorylated-P38, anti-P38, anti-phosphorylated-ERK, anti-ERK, anti-cleaved-caspase-3, and anti-GAPDH (Cell Signalling Technology, Danvers, MA, United States).
5
Protein Expression Analysis by Western Blot
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RIPA buffer containing a protease inhibitor (Beyotime, Nantong, China) was used to lyse the tissues or cells. Equal amounts of protein were separated using 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (NCM Biotech, Suzhou, China) and then transferred onto nitrocellulose filter membranes (Millipore, USA). The membranes were blocked with 5% skim milk for 2 h. After washing twice with Tris-Buffered Saline Tween-20 buffer (Sevicebio, Wuhan, China), the membranes were incubated with primary antibodies, including anti-Fatty Acid Synthase (1:1000; Cell Signaling Technology), anti-GAPDH (1:5000; Proteintech), anti-xCT (1:1000; Cell Signaling Technology), anti-GPX4 (1:1000; Cell Signaling Technology), anti-CD71 (1:1000; Cell Signaling Technology), anti-p-ERK (1:1000; Cell Signaling Technology), or anti-ERK (1:1000; Cell Signaling Technology) overnight, followed by anti-rabbit IgG (1:2000; Cell Signaling Technology) or anti-mouse IgG (1:5000; Abmart) for 2 h. Lastly, the images were developed with ChemiScope 6100 (Clinx, Shanghai, China) after adding chemiluminescent substrate (NCM biotech).
6
Western Blot Analysis of Lipogenic Enzymes
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Total proteins of cell lysate or liver homogenates were separated on SDS-PAGE, then transferred to polyvinylidene fluoride membranes (Millipore, Billerica, MA, USA). The antibodies used in western blot assay were anti-Acetyl-CoA Carboxylase (ACC); anti-Fatty Acid Synthase (FAS), and anti-SCD1 antibodies (Cell Signaling Technology, 1:1000); anti-Mcad, anti-LCAD and anti- SCAD antibodies (Abcam, 1:1000). The immune complexes were detected by enhanced chemiluminescence (Millipore, Billerica, MA, USA). Anti-GAPDH (Beyotime, 1:1000) was as an internal control. The band was quantified using Image Lab software (BioRad laboratories, Hercules, CA, USA). Each experiment was performed at least twice.
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