Islet proteins (∼150) lysed in RIPA buffer with protease inhibitors (Millipore) were separated by SDS-PAGE and blotted with anti-PGC-1α (1:1000, Calbiochem 4Cl.3), anti-ATGL (1:1000, Cell Signaling #2138) and anti-HSL (1:1000, Cell Signaling #4107).
Anti atgl
Manufactured by Cell Signaling Technology
Sourced in United States
Anti-ATGL is a primary antibody that recognizes the Adipose Triglyceride Lipase (ATGL) protein. ATGL is an enzyme involved in the hydrolysis of triglycerides, playing a crucial role in lipid metabolism. The Anti-ATGL antibody can be used to detect and quantify ATGL expression in various cell and tissue samples.
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Lab products found in correlation
Anti hsl, by Cell Signaling Technology (8 mentions)
Insulin, by Merck Group (3 mentions)
Anti ampk, by Cell Signaling Technology (3 mentions)
Anti gapdh, by Cell Signaling Technology (2 mentions)
Anti β actin, by Cell Signaling Technology (2 mentions)
Anti glut4, by Cell Signaling Technology (2 mentions)
Protease inhibitor, by Merck Group (1 mentions)
Pvdf membrane, by Merck Group (1 mentions)
Anti cpt1a, by Abcam (1 mentions)
Total oxphos complex kit, by Thermo Fisher Scientific (1 mentions)
Anti α sma, by Thermo Fisher Scientific (1 mentions)
Chemidoc touch imaging system, by Bio-Rad (1 mentions)
Ecl plus western blotting detection reagent, by Thermo Fisher Scientific (1 mentions)
Polyvinylidene fluoride membrane, by Carl Roth (1 mentions)
Anti calnexin, by Cell Signaling Technology (1 mentions)
3 isobutyl 1 methylxanthine, by Merck Group (1 mentions)
Anti flag m2 agarose, by Merck Group (1 mentions)
Fugene hd, by Promega (1 mentions)
Oleic acid, by Merck Group (1 mentions)
Protein g agarose, by Thermo Fisher Scientific (1 mentions)
19 protocols using anti atgl
1
Islet Protein Expression Analysis
2
Immunoblotting of Cell Lysates
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Cell lysates were prepared as previously described [28 (link)]. Cell lysates were subjected to SDS-PAGE, transferred to PVDF membranes (Merck Millipore), and then immunoblotted with antibodies for anti-ATGL (#2138), anti-HSL (#4107), and anti-GAPDH (#2118) obtained from Cell Signaling Technology (Danvers, MA), Total OxPhos Complex Kit (# 458099) from Invitrogen (Life Technologies Australia Pty Ltd), anti-14-3-3 (sc-33752) from Santa Cruz Biotech (Dallas, TX), and anti-CPT1A (#ab128568) from Abcam (Cambridge, MA).
3
Immunoblotting Analysis of Protein Expression
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Analysis of protein expression by immunoblotting was performed as previously described (10 (link)). Proteins of cell lysates (20–40 μg) were dissolved in SDS sample buffer, separated by 10% or 12.5% SDS-PAGE, and transferred onto a polyvinylidene fluoride membrane (Carl Roth GmbH, Karlsruhe, Germany). The membrane was blocked with 10% nonfat dry milk and incubated with the following primary antibodies: anti-ATGL, anti-HSL, anti-GAPDH, and anti-calnexin from Cell Signaling Technology (Danvers, MA; 2138S/ATGL, 4107S/HSL, 2118S/GAPDH, 2679S/calnexin), anti-6x HIS tag, and anti-NADH:ubiquinone oxidoreductase core subunit S1 (NDUFS1) from Abcam (Cambridge, England; ab18184/6x HIS-tag®, ab157221/NDUFS1), anti-α-SMA from Thermo Fisher Scientific (Waltham, MA; PA5-22251/α-SMA), and anti-KIAA1363 from Invitrogen GmbH (PA5-50285/NCEH1 = KIAA1363), respectively. For detection, membranes were incubated with horseradish peroxidase-labeled secondary antibodies specific for respective primary antibody. Bands were visualized using the ECL plus Western blotting Detection Reagent (Thermo Fisher Scientific) and ChemiDoc Touch Imaging System (Bio-Rad).
4
Characterization of LDAH Antibodies
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Custom polyclonal rabbit anti-human and mouse LDAH antibodies were generated at Bethyl Laboratories, and characterized as previously described15 (link). Bodipy 493/503, LipidTox, and RNAiMAX were from Invitrogen. Fugene HD was purchased from Promega. Isoproterenol hydrochloride, anti-beta actin antibody, anti Flag-M2-HRP, anti Flag-M2-agarose, mouse IgG, oleic acid, 3-isobutyl-1-methylxanthine, insulin, dexamethasone and nocodazole were from Sigma-Aldrich. Triacsin C and anti-ubiquitin antibody were from Santa Cruz Biotechnology. Anti-PLIN1 antibody was from Progen. Anti-beta tubulin and anti-PLIN2 antibodies were from Nobus Biologicals. Anti-PLIN3 was purchased from Fitzgerald. Anti-ATGL and anti-HSL were from Cell Signaling. Alexa Fluor 647-conjugated secondary antibodies and anti-GM130 were from BD Pharmingen. Protein G-agarose and protein-A-agarose were from Thermo Scientific. MG132 was purchased from Calbiochem. Total and free cholesterol were measured using kits from Wako. TAG were determined with Infinity Trglyceride reagents from Thermo Scientific. 14C-oleic acid was purchased from Perkin Elmer.
5
Adipose Tissue Protein Analysis
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The following antibodies were purchased from Cell Signaling Technology: anti-ATGL (#2439, 1:1000, RRID:AB_2167953), anti-Phospho-HSL (#4137,1:1000, RRID:AB_2135498), anti-HSL (#4107,1:1000, RRID:AB_2296900), and anti-Phospho-PKA (#9621,1:1000, RRID:AB_330304). The following antibodies were purchased from Abcam: anti-cAMP (ab76238,1:1000, RRID:AB_1523259) and anti-UCP1 (ab234430,1:1000, RRID:AB_2905638).
6
Antibody Panel for Adipogenesis Signaling
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The following antibodies were used in this study: anti-GAPDH (sc-25778; Santa Cruz Biotechnology, Dallas, TX); anti-Pik3r1 (4292s; Cell Signaling Technology, Danvers, MA), anti-ATGL (2138s; Cell Signaling Technology); anti-Akt (9272s; Cell Signaling Technology); anti–phospho-Akt (T308) (9275s; Cell Signaling Technology); anti–perilipin A (ab3526; Abcam, Cambridge, MA); anti–phospho-perilipin (4856; VALA Sciences, San Diego, CA); anti-HSL (4107s; Cell Signaling Technology); anti–phospho-HSL (S660) (4126s; Cell Signaling Technology); anti–PKA-RIα (610609; BD Biosciences, San Jose, CA); anti–PKA-RIIβ (610625; BD Biosciences); and anti-PKAα catalytic subunit (C-20) (sc-903; Santa Cruz Biotechnology). Anti-Ubxd8 antibody was provided by the laboratory of Dr. James Olzmann (University of California, Berkeley). The intensity of the bands was quantified using ImageJ software (National Institute of Health) and normalized to GAPDH or Ubxd8 as indicated.
7
Antibodies for Poliovirus and Host Factors
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Mouse monoclonal anti-poliovirus 2C and 2B were described in [105 (link)]. Rabbit polyclonal anti-polio 3D antibodies were developed by Chemicon using recombinant 3D protein as immunogen. Rabbit monoclonal anti-CCTα, anti-eIF2α, anti-eIF2α (phospho Ser51), anti-IRF3, anti-IRF3 (phospho Ser396), anti-HSL, anti-ATGL, anti-STAT1, anti-Viperin, anti-ISG15 and anti-HA antibodies used in western blots and immunofluorescence were from Cell Signaling. Mouse monoclonal anti-dsRNA antibodies were from English and Scientific Consulting Kft. Mouse monoclonal anti-GBF1 antibodies were from BD Biosciences; rabbit anti-PI4KIIIβ were from EMD Millipore. Mouse monoclonal antibodies against Iκβ were a kind gift from Dr. John Patton (University of Maryland). Alexa dyes conjugated antibodies were from Molecular Probes (Thermo Fisher); secondary HRP-conjugated antibodies were from Amersham. Mouse monoclonal anti-HA antibody used for co-IP was from Santa Cruz Biotechnology.
8
Western Blot Analysis of Adipose Tissue Proteins
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Expression of HSL, its serine-563-phosphorylated form (HSL Ser563) and ATGL were measured in epididymal fat pad homogenates by western blot. Antibodies for immunoblotting were as follows: anti-HSL (1:2000, Cell Signaling Technology, USA), anti-HSL Ser563 (1:500, Cell Signaling Technology), anti-ATGL (1:2000, Cell Signaling Technology) and anti-β-actin (1:5000, Sigma). Epididymal adipose tissue homogenates (25 µg total protein per well) were separated by SDS page, and proteins transferred to a polyvinylidene fluoride membrane. Membranes were then blocked [5% milk and 0.1% TWEEN-20 in PBS (PBS-T)], and incubated at 4°C overnight in primary antibodies diluted in 1% BSA in PBS-T. After being incubated with HRP-conjugated secondary antibodies (1:500, Wako Chemicals) in 5% milk in PBS-T, blots were developed using an Enhanced Chemiluminescent Detection Kit (GE Healthcare). Digital images of protein bands were acquired (ImageQuant 350, GE Healthcare, UK) and semi-quantitative analysis of protein content performed by densitometry using ImageQuant TL software (GE Healthcare, UK), with values normalised to β-actin as a loading control.
9
Western Blot Analysis of Adipogenic Proteins
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Western blot analysis was conducted as previously reported.15 (link) The target protein was detected using primary antibodies as follows: anti-proliferator-activated receptor gamma (PPAR-γ) (1:1000, Cell Signaling Technology), anti-PGC-1α (1:1000, Abcam), anti-ATGL (1:1000, Cell Signaling Technology), anti-pHSL (1:1000, Cell Signaling Technology), anti-tHSL (1:1000, Cell Signaling Technology), anti-α-tubulin (1:1000, Abcam) and anti-β-actin (1:2000, Cell Signaling Technology).
10
Immunohistochemical Analysis of Lipid Metabolism
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