Abl (ABL1) (Q252H) Human TK Kinase Enzymatic Radiometric [Km ATP] KinaseProfiler LeadHunter Assay - FR

Item: 14-751KP

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OVERVIEW

DOCUMENTS/REFERENCES

Biological Information

Background Information:

The abl oncogene was initially identified as the viral transforming gene (v-abl) of Abelson murine leukemia virus (A-MuLV), from a chemically thymectomized mouse inoculated with Moloney murine leukemia virus (M-MuLV). The v-abl genome consists of M-MuLV gag sequences fused to mouse c-abl, resulting in the production of phosphoroteins, p160 and p120 gag/c-abl with protein-tyrosine kinase activity, as the major translational products. The abl oncogene is implicated in serval human leukemias including chronic myelocytic (CML), adult acute lymphoblastic (ALL) and pediatric ALL. In these leukemias the c-abl proto-oncogene undergoes a chromosomal translocation producing the Philadelphia (Ph¢) chromosome resulting in the fusion of c-abl sequences to the bcr gene of chromosome 22, and the molecular consequences of this translocation is the generation of a chimeric bcr/c-abl mRNA encoding activated abl protein-tyrosine kinase.

Family:

Kinase

Sub Family:

Class:

Protein Tyrosine

Protein Name:

Abl

Uniprot Number:

P00519

Protein Aliases:

Abelson murine leukemia viral oncogene homolog 1|Abelson tyrosine-protein kinase 1|Proto-oncogene c-Abl|p150

Gene Name:

ABL1

Gene ID:

Gene Aliases:

JTK7|c-ABL|p150

Accession Number:

U07563

Organism:

Human

Construct Details:

27-end; Q252H recombinant

Assay Information

Assay Type:

Biochemical

Assay Sub Type:

Enzymatic

Functional Mode:

Antagonist

Detection Method:

Radiometric

Measured Response:

Scintillation

Testing Information

Procedure Summary:

Abl (Q252H) (h) is incubated with 8 mM MOPS pH 7.0, 0.2 mM EDTA, 50 µM EAIYAAPFAKKK, 10 mM MgAcetate and [g- 33P]-ATP (specific activity and concentration as required). The reaction is initiated by the addition of the Mg/ATP mix. After incubation for 40 minutes at room temperature, the reaction is stopped by the addition of phosphoric acid to a concentration of 0.5%. An aliquot of the reaction is then spotted onto a filter and washed four times for 4 minutes in 0.425% phosphoric acid and once in methanol prior to drying and scintillation counting.

Substrate:

50 uM EAIYAAPFAKKK

Tracer:

33P

ATP Concentration:

[ATP]= 15µM (Km= 26.5µM )

Incubation:

40 min at Room temperature

Control Inhibitor:

Staurosporine

Test Sample Requirements:

Please provide compounds dissolved in 100% DMSO (PBS or HBSS prohibited).
Stock solution should be ≥50× the final assay concentration (a 10 mM stock is also acceptable).
Required volume:
– <60 kinase assays: 120 µL of 50× stock
– ≥60 assays: 420 µL of 50× stock
If you supply 10 mM stocks or powder (pre-weighed), we will apply the appropriate conversion.
For batches of ~50 compounds supplied as solids, the TAT will be extended.

Minimum Order Quantity:

Turnaround Time

Standard:

6 Business Days

Service Scheduling Note:

Assay is run on a weekly schedule beginning on Friday. Compounds and purchase order must be received on or before 13:00 CET Thursday for the order to be included on the service run that begins on Friday of the same week.

Reference Compound Data

PubChem ID

122130844

Name

Staurosporine

Measurement

36.33nM - IC50

Clinical Relevance

Therapeutic Area:

Oncology/Immuno-Oncology

Additional Information

Brand:

LeadHunter

Testing Location:

KP - France - Celle L'Evescault