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Encompassing Amino Acids: 498-796
Applications: Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.
Assay Conditions: The activity of the complex was measured using peptide sequence (EAIYAAPFAKKK), based on the C-terminus of Abl, as a substrate (stock solution 1 mg/ml dissolved in distilled water).
Assay: Increasing amounts of kinase were mixed with a final concentration of 200 µg/ml peptide substrate in a buffer consisting of 15 mM MOPS pH 7.2, 15 mM MgCl2, 7.5 mM β-glycerol-phosphate, 3 mM EGTA, 1.2 mM EDTA, 20 µg/ml BSA (bovine serum albumin) and 150 µM fresh DTT (final concentrations). The reaction was initiated by adding a [33P]-ATP (1 µCi/sample) mixed with non-radioactive ATP to reach a final concentration of 50 µM. The blank was determined from a "no substrate" sample.
The reaction was incubated for 15min at 30°C and terminated by spotting 20 µl of the mixture onto phosphocellulose paper strips that were fixed in 1% phosphoric acid and washed three times. Radioactivity was determined using a scintillation counter.
Description: Recombinant human LTK (leukocyte receptor tyrosine kinase, also known as TYK1), encompassing amino acids 498-796 containing the protein kinase domain. This construct contains an N-terminal GST-tag. The recombinant protein was affinity purified and is kinase-active.
Format: Aqueous buffer solution
Formulation: 50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 10 mM glutathione, 0.1 mM EDTA, 0.25 mM DTT, 0.1 PMSF, 25% glycerol
Genbank: NM_002344
Purity: ≥90%
Storage Stability: At least 6 months at -80°C.
Tags: N-terminal GST-tag
Uniprot: P29376
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
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