ALK (T1151 L1152insT), GST-Tag Recombinant

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Encompassing Amino Acids: 1060-end

Applications: Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.

Assay Conditions: ALK (T1151 L1152insT) activity was measured by using the IGF1Rtide synthetic peptide (KKKSPGEYVNIEFG) diluted in 20 mM Tris-HCl, pH 7.5 to a working concentration of 1 mg/ml, in the ADP Glo™ Kinase Assay kit (Promega #V9101). Reaction was initiated by mixing increasing amounts of ALK (T1151 L1152insT) with 25 µM ATP in 40 mM Tris-HCl, pH 7.4, 20 mM MgCl₂, 2.5 mM MnCl₂, 0.1 mg/ml BSA prepared with 50 µM DTT and substrate at a final concentration of 200 µg/ml.After a 40-minute incubation at room temperature, the reaction was terminated by addition of ADP-Glo™ Reagent, followed by a subsequent 40-minute incubation at room temperature. Kinase Detection Reagent was added, and the reaction was incubated for another 30 minutes at ambient temperature. Detection of luminescence was measured using the Luminescence Module Protocol on GloMax®-Multi Microplate Multimode Reader. The Specific Activity was calculated as follows: (Corrected activity, RLU) / [(Specific activity from ADP in RLU/pmol) * (Reaction time in min) *(Enzyme amount in µg or mg)].  Corrected RLU was calculated by subtracting the blank value from all the values. The blank was determined from a "no enzyme" sample by replacing the enzyme solution with an equal volume of Dilution Buffer IX (1x).

Background: ALK (anasplatic lymphoma kinase), also known as ALK tyrosine kinase receptor or CD246 (cluster of differentiation 246), is a receptor tyrosine kinase involved in signal transduction. In the presence of a ligand ALK dimerizes and a conformational change result in autoactivation of the kinase domain. Activated ALK will phosphorylate other ALK receptors and activate downstream signaling pathways. ALK is present in the nervous system during development, where it participates in retinal axon growth and targeting, synapse development, sleep, learning and long-term memory. Interestingly, dysfunction of ALK in one of three possible ways can lead to cancer: fusion with another gene, gene duplication or gene mutations. ALK, as its name indicates, has been linked to anaplastic large-cell lymphoma, but also non-small-cell lung cancer (NSCLC), neuroblastoma, breast cancer, renal carcinoma and others. Inhibitors of ALK show great therapeutical potential, two of them being already commercially available for the treatment of late-stage lung cancer and NSCLC. Further studies into ALK will deepen our understanding of its functions, find new inhibitors and new therapeutic avenues for patients with ALK-linked cancer.

Description: Recombinant human ALK (T1151 L1152insT) (anaplastic lymphoma kinase), encompassing amino acids 1060-end with the T1151 L1152insT mutations. This construct contains an N-terminal GST-tag. The recombinant protein was affinity purified and is 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 mM PMSF, 25% glycerol

Genbank: NM_004304

Purity: ≥70% (Purity calculation does not include co-purifying Glutathione-binding proteins.)

Storage Stability: At least 6 months at -80°C.

Tags: N-terminal GST-tag

Uniprot: Q9UM73

Warnings: Avoid freeze/thaw cycles

Biosafety Level: Not applicable (BSL-1)

• Artikelnummer: BPS101930
• Hersteller: BPS Bioscience
• Hersteller Artikelnummer: 101930
• Green Labware: Nein
• Verpackungseinheit: 10 µg
• Mengeneinheit: STK
• Wirt: Human