Commercially available chemicals were used as received. All reagents and Sephadex ion-exchange resin were obtained from Sigma-Aldrich with the following exceptions. RhCl3 was purchased from Pressure Chemical, Inc. Dowex ion-exchange beads were purchased from Acros Organics. Analytical standards for Rb and transition metals were purchased from Analytical West and Ultra Scientific, respectively. MTT and ELISA assay kits were obtained from Roche. Pierce BCA assay kit and NP40 were purchased from Thermo Scientific. Sep-pak C18 solid-phase extraction (SPE) cartridges were purchased from Waters Chemical Co. Cell culture media and supplements were purchased from Invitrogen. Tissue culture flasks and plates were obtained from Corning. 32P labeled ATP was purchased from Perkin Elmer. UreaGel supplies were purchased from National Diagnostics. Microbiospin columns were purchased from BioRad.
32p labeled atp
Manufactured by PerkinElmer
Sourced in United States
32P-labeled ATP is a radioactive compound used in various biochemical and molecular biology applications. It is produced by incorporating the radioactive isotope phosphorus-32 (32P) into the phosphate groups of adenosine triphosphate (ATP) molecules. This product can be used as a tracer or substrate in experimental procedures where the detection or quantification of ATP-dependent processes is required.
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Lab products found in correlation
Pierce bca assay kit, by Thermo Fisher Scientific (1 mentions)
Np 40, by Thermo Fisher Scientific (1 mentions)
Cell culture media and supplements, by Thermo Fisher Scientific (1 mentions)
Tissue culture flasks and plates, by Corning (1 mentions)
Micro bio spin column, by Bio-Rad (1 mentions)
T4 polynucleotide kinase, by Merck Group (1 mentions)
Actin fs activated ptt reagent, by Siemens (1 mentions)
Test thrombin reagent, by Siemens (1 mentions)
Thromborel s, by Siemens (1 mentions)
M116487001, by Merck Group (1 mentions)
Typhoon fla 7000, by GE Healthcare (1 mentions)
Imagequant, by GE Healthcare (1 mentions)
Assay buffer 1, by Sino Biological (1 mentions)
4 protocols using 32p labeled atp
1
Comprehensive Reagent Procurement for Multidisciplinary Research
2
Aptamer-Based Anticoagulant Formulation
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DNA aptamer RA-36, dGGTTGGTGTGGTTGGTGGTTGGTGTGGTTGG⋅2 KCl, was synthesized and purified up to pharmaceutical grade for intravenous injections at ‘APTO-PHARM,’ Ltd. Russian Federation. The drug formulation for intravenous injections contained 10 mg/ml of the active substance in 0.9% sodium chloride solution; to prepare that formulation, the aptamer stock solution was diluted with 0.9% sodium chloride immediately before the intravenous injection, when proper dosing is needed.
Lyophilized bivalirudin (Angiox®, lot# PL2097) was purchased from Medicines Company, United Kingdom. The powder was dissolved in 0.9% sodium chloride at a concentration of 5 mg/ml according to the manufacturer’s guidelines. The stock solution was diluted with 0.9% sodium chloride immediately before intravenous injection, when proper dosing is needed.
Standard human plasma, Test Thrombin Reagent, Thromborel® S and Actin FS Activated PTT Reagent were purchased from Siemens, Germany. 32P-labeled ATP with beta radiation activity of 20 MBq was purchased from Perkin-Elmer, United States. T4 polynucleotide kinase (Sigma–Aldrich, United States).
Lyophilized bivalirudin (Angiox®, lot# PL2097) was purchased from Medicines Company, United Kingdom. The powder was dissolved in 0.9% sodium chloride at a concentration of 5 mg/ml according to the manufacturer’s guidelines. The stock solution was diluted with 0.9% sodium chloride immediately before intravenous injection, when proper dosing is needed.
Standard human plasma, Test Thrombin Reagent, Thromborel® S and Actin FS Activated PTT Reagent were purchased from Siemens, Germany. 32P-labeled ATP with beta radiation activity of 20 MBq was purchased from Perkin-Elmer, United States. T4 polynucleotide kinase (Sigma–Aldrich, United States).
3
MBP-LecRK-I.8KD Kinase Assay
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The kinase assay was performed as described by Choi et al. (2014) (link) with minor modifications. Briefly, 5 µg of MBP-LecRK-I.8KD and 2 µg of myelin basic protein in a buffer (50 mM Tris-HCl pH7.5, 50 mM KCl, 10 mM MnCl2, 10 mM MgCl2) plus 1 µL of ATP (0.4 µL 32P-labeled ATP, 0.4 µL cold ATP, 0.2 µL H2O) in a total volume of 30 µL were incubated at 30°C for 30 min. 32P-labeled ATP (specific activity 3000 Ci/mmol) was purchased from PerkinElmer. After SDS-PAGE electrophoresis, the gel was dried and exposed to X-ray film for 3 hr.
4
PIP2 Phosphorylation Assay Protocol
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For phosphorylation reactions, predetermined concentrations of protein kinase and PI3K complexes were added to master mix containing 50 mM of ATP, 0.1 mg/mL of BSA and 1x Assay Buffer I (SignalChem) in 40 mL total volumes. Reactions were carried out at 30°C for 30 min. Next, 0.01 mCi/mL 32P-labeled ATP (PerkinElmer) and 25 μM PIP2 (Thermo Scientific) were included, bringing total volume to 100 mL PIP2 phosphorylation reactions were carried out at 30°C for 20 min 50 μl of 4N HCl was added to quench the reaction and the lipid was extracted with 100 mL of CHCl3: MeOH (1:1) followed by vortexing for 1 min and centrifugation at maximum speed for 2 min. 10 mL of the lower hydrophobic phase was extracted with gel loading pipet tips and spotted onto a silica plate (EMD Millipore #M116487001) for thin-layer chromatography. Plates were placed in a sealed chamber with 1-propanol: 2M acetic acid (65:35). Radioactivity was imaged with a Typhoon FLA 7000 phosphorimager (GE) and quantified by ImageQuant (GE). For cationic liposomes, EPC (#890704C) and PI (840042C) were purchased from Avanti. Liposomes containing 5% PI, 60% EPC, and 35% cholesterol were prepared by extrusion via 20 passes through a 0.1 mm membrane using a Mini-Extruder kit (Avanti). The anionic liposomes in these experiments replaced EPC with PS.
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