Synthetic dioleolyl phospholipids PS (phosphatidylserine; 1,2-dioleoyl-sn-glycero-3-phospho-L-serine), PIP2 (1,2-dioleoyl-sn-glycero-3-phosphoinositol 4,5-diphosphate), and PE (1,2-dioleoyl-sn-glycero-3-phosphoethanolamine) were from Avanti Polar Lipids (Alabaster, AL). Alexa Fluor 555 C2-maleimide (AF555) and CoverWell perfusion chambers were from Invitrogen (Carlsbad, CA). Glass supports were from Ted Pella, Inc. (Redding, CA). 2-Mercaptoethanol, ultrapure (>99%) BSA, ATP magnesium salt, and CoA trilithium salt were from Sigma (St. Louis, MO). Monophosphorylated peptide (pY) (sequence of RQIKIWFQNRRMKWKKSDGGpYMDMS) produced by TOCRIS Bioscience (Bristol, U.K.) contained the conserved sequence of the PDGFR Y740 phosphorylation site (SDGGpYMDMS). This peptide activated PI3Ka to the same level as the bis-phosphorylated PDGFR-derived peptide we used previously (compare the results presented here with those in ref 26 (link) and data not shown). Ultrapure (≥99%) CHAPS {3-[(3-cholamidopropyl)-dimethylammonio]-1-propanesulfonate} was from Anatrace (Maumee, OH). Human MARCKS PIP2 binding domain (MARCKS residues 151–175) was produced by SynBioSci Corp. (Livermore, CA) and includes an N-terminal cysteine residue added for probe labeling (N-CKKKKKRFSFKKSFKL-SGFSFKKNKK-C).
Atp magnesium salt
Manufactured by Merck Group
Sourced in Germany
ATP magnesium salt is a laboratory reagent that provides a source of adenosine triphosphate (ATP) and magnesium ions. ATP is a fundamental energy-carrying molecule in living cells, and magnesium is a cofactor required for many enzymatic reactions. This product is typically used in biochemical and cell biology research applications where a stable source of ATP and magnesium is needed.
Automatically generated - may contain errors
Lab products found in correlation
Supersome, by Corning (2 mentions)
Methanol, by Avantor (2 mentions)
Ammonium acetate, by Merck Group (2 mentions)
Sodium orthovanadate, by Merck Group (2 mentions)
Sulfasalazine, by Merck Group (2 mentions)
Mes hydrate, by Merck Group (2 mentions)
Coverwell perfusion chamber, by Thermo Fisher Scientific (1 mentions)
2 mercaptoethanol, by Merck Group (1 mentions)
Coa trilithium salt, by Merck Group (1 mentions)
Acetylcholine chloride, by Merck Group (1 mentions)
Sodium bicarbonate, by Merck Group (1 mentions)
Sodium chloride (nacl), by Merck Group (1 mentions)
Sucrose, by Merck Group (1 mentions)
Ethylene glycol tetraacetic acid (egta), by Merck Group (1 mentions)
Hepes, by Merck Group (1 mentions)
Ethylene diamine tetraacetic acid (edta), by Merck Group (1 mentions)
Polyethyleneimine, by Merck Group (1 mentions)
Magnesium sulfate, by Merck Group (1 mentions)
L tartaric acid, by Merck Group (1 mentions)
L ascorbic acid, by Merck Group (1 mentions)
6 protocols using atp magnesium salt
1
Phospholipid Reconstitution for Signaling
2
Stinging Nettle Extracts Pharmacological Analysis
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Stinging nettle root and leaf extracts (500 mg dried plant material/ml in 25% alcohol) were purchased commercially from MediHerb® Pty. Ltd. (Warwick, Queensland, Australia). Certified reference plant material was checked against the British Pharmacopoeia and authenticated by Mr Howard Hollow, Southern Cross University, Centre for Phytochemistry and Pharmacology (Lismore, NSW, Australia). The plant voucher specimen (reference material: MediHerb batch number T7C068; sample ID #CPA080318) is retained at Southern Cross University. These extracts are available in Australia for practitioner dispensing. Noradrenaline bitartrate salt (Arterenol®, Sigma) was dissolved and diluted to required concentrations using a catecholamine diluent (NaCl 154 mM, NaH2PO4 1.2 mM, ascorbic acid 0.2 mM in distilled water). Acetylcholine chloride (Sigma), ATP magnesium salt (Sigma), and αβ-methylene ATP lithium salt (Sigma) were all dissolved and diluted to required concentrations using distilled water. All drugs were made fresh on the morning of experimentation. Water used in all experiments was distilled using the MilliPore system (Burlington, MA, U.S.A.).
3
Radiolabeled Dopamine Binding Assay
4
Purification and Characterization of hMAGL
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Reactions progress was monitored by analytical thin-layer chromatography (TLC) on precoated aluminum foil (Silica Gel 60 F254-plate, Sigma–Aldrich, St. Louis, MO, USA), and the products were visualized by UV light. Purity of all compounds (>98%) was verified by thin layer chromatography and NMR measurements [46 (link),47 (link)]. The chemicals required for validation, all of analytical grade, and all reagents required for bacterial cell culture and the kit for plasmid extraction were purchased from Sigma–Aldrich (St. Louis, MO). E. coli JM109 competent cells were from Promega, (Fitchburg, WI, USA). Ni-NTA agarose and 3–12 mL 10kDa Slide-A-Lyzer™ Dialysis Cassette used for protein purification were purchased from Thermo Fisher Scientific (Waltham, MA, USA). SDS-PAGE experiments were performed using Mini-PROTEAN® II handcast systems (Bio-Rad, Hercules, CA, USA) kits for both equipment and reagents, unless specified. Monoacylglycerol lipase (human recombinant, 50 μg, hMAGL) was purchased from Cayman Chemical (Ann Arbor, MI, USA). For bioluminescence experiments, LH2 potassium salt was purchased from Promega Italia Srl, while ATP magnesium salt was from Sigma–Aldrich.
5
Baculovirus-Expressed BCRP Transporter Assay
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The baculovirus-infected insect cell microsomes (Supersomes) containing human complementary DNA-expressed BCRP (Arg482, 5 mg protein/mL) and wildtype Supersomes without hBCRP (control membrane, 5 mg protein/mL) used as negative control were obtained from Corning (Amsterdam, The Netherlands). After delivery, Supersomes were thawed at 37 °C, aliquoted, snap-frozen in liquid nitrogen, and stored at -80 °C until use.
AMP disodium salt, ADP sodium salt, ATP magnesium salt, guanosine 5'diphospate (GDP) sodium salt, uridine 5'-phosphate (UDP) sodium salt hydrate, sulfasalazine, sodium orthovanadate, amprenavir, indinavir, nelfinavir, ritonavir, saquinavir mesylate, ammonium acetate, MES hydrate, and Trizma base were obtained from Sigma-Aldrich (Taufkirchen, Germany), formic acid (MS grade) from Fluka (Neu-Ulm, Germany), acetonitrile, methanol (both LC-MS grade), and all other chemicals from VWR (Darmstadt, Germany).
Stock solutions were prepared in bidistilled water for sodium orthovanadate (10 mM), AMP, ADP, ATP, GDP, and UDP (20 mM, respectively) or in methanol for sulfasalazine (0.5 mg/mL), amprenavir, indinavir, nelfinavir, ritonavir, and saquinavir (1 mg/mL, respectively). Stock solutions were aliquoted and stored at -20 °C until use.
AMP disodium salt, ADP sodium salt, ATP magnesium salt, guanosine 5'diphospate (GDP) sodium salt, uridine 5'-phosphate (UDP) sodium salt hydrate, sulfasalazine, sodium orthovanadate, amprenavir, indinavir, nelfinavir, ritonavir, saquinavir mesylate, ammonium acetate, MES hydrate, and Trizma base were obtained from Sigma-Aldrich (Taufkirchen, Germany), formic acid (MS grade) from Fluka (Neu-Ulm, Germany), acetonitrile, methanol (both LC-MS grade), and all other chemicals from VWR (Darmstadt, Germany).
Stock solutions were prepared in bidistilled water for sodium orthovanadate (10 mM), AMP, ADP, ATP, GDP, and UDP (20 mM, respectively) or in methanol for sulfasalazine (0.5 mg/mL), amprenavir, indinavir, nelfinavir, ritonavir, and saquinavir (1 mg/mL, respectively). Stock solutions were aliquoted and stored at -20 °C until use.
6
Baculovirus-Insect Cell Microsomes for BCRP
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Baculovirus-infected insect cell microsomes (Supersomes) containing complementary DNA-expressed hBCRP (Arg482, 5 mg protein/mL) and wild-type Supersomes without hBCRP (control membrane, 5 mg protein/mL) used as negative control were obtained from Corning (Amsterdam, The Netherlands). After delivery, Supersomes were thawed at 37 °C, aliquoted, snap-frozen in liquid nitrogen, and stored at -80 °C until use. ADP sodium salt, ATP magnesium salt, sulfasalazine, rosuvastatin, sodium orthovanadate, ammonium acetate, MES hydrate, and TRIS base were obtained from Sigma-Aldrich (Taufkirchen, Germany), formic acid (MS grade) from Fluka (Neu-Ulm, Germany), acetonitrile, methanol (both LC-MS grade), and all other chemicals from VWR (Darmstadt, Germany). -ethyl-8-methoxy-1,2,3,4,6,7,12,12boctahydroindolo[2,3-a] quinolizin-2-yl]-3-methoxyprop-2-enoate (mitragynine) by the Department of Forensic Medicine, Johannes Gutenberg University (Mainz, Germany), where it was isolated from kratom leaves obtained from head&nature (Regensburg, Germany) (Philipp et al. 2009) . N-Allyl-N- [2-(5-methoxy-1H-indol-3-yl) ethyl]-2-propen-1-amine (5-MeO-DALT) was synthesized by use of established methods (Brandt et al. 2008 ) and provided by the School of Pharmacy and Biomolecular Sciences, John Moores University (Liverpool, UK).
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