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7405
HTScan® Profiling Kit (Tyrosine Kinase Set I)
Cellular Assay Kits

HTScan® Profiling Kit (Tyrosine Kinase Set I) #7405

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Compound panel (5 µM each compound): DELFIA® data were generated using HTScan® Profiling Kit (Tyrosine Kinase Set I) to detect the inhibition of various RTKs by a panel of compounds (Staurosporine, Iressa, SU-6668 and CT53518). In a 50 µl reaction, 10 Units of each RTK, 1.5 uM substrate peptide, 20 uM ATP and 5 uM inhibitor were used per reaction well (in duplicate) at 25ºC for 30 minutes. (DELFIA® is a registered trademark of PerkinElmer, Inc.)

Dose response (SU-6668): DELFIA® data were generated using HTScan® Profiling Kit (Tyrosine Kinase Set I) to detect the inhibition profile of SU-6668 toward various RTKs. In a 50 ul reaction, 10 Units of each GST-RTK, 1.5 uM substrate peptide, 20 uM ATP and various concentrations of SU6668 were used per reaction well at 25ºC for 30 minutes. (DELFIA® is a registered trademark of PerkinElmer, Inc.)

Assay plate (96 well format): Peptide substrates for the indicated eight RTKs have been arrayed in 96-well format.

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Product Description

The kit provides a means of characterizing the effect of a compound or panel of compounds on the included active human receptor tyrosine kinases.

The tyrosine kinases in the kit are supplied as GST fusion proteins. Biotinylated substrate peptides validated for each kinase are supplied at 6µM, and a phosphotyrosine monoclonal antibody for detection of the phosphorylated form of the substrate peptide is also included in the kit. The kit enables rapid characterization of the effect of a single compound at multiple concentrations or a panel of compounds toward the following RTKs: PDGFR-beta, EGFR, FGFR-3, VEGFR-2, FLT3, IGF1R, MET and EPHB3, each representing an RTK subgroup.

Specificity / Sensitivity

The purity of each RTK was characterized using SDS-PAGE, and assay conditions for phosphorylation of the included substrates were optimalized using DELFIA® (for details, visit www.cellsignal.com/drug2.AS) (DELFIA® is a registered trademark of PerkinElmer, Inc.).

Background

Tyrosine kinases have been validated as targets for therapeutic intervention in many disease areas. The human genome codes for nearly one hundred tyrosine kinases, which can be categorized into receptor tyrosine kinases (RTKs) and cytosolic tyrosine kinase. RTKs are currently a major focus for therapeutic drug target screening in the pharmaceutical industry. HTScan¬Profiling Kit (Tyrosine Kinase Set 1) from Cell Signaling Technology (CST) is an antibody-based kinase activity assay kit.

Each kinase is paired with a validated biotynilated substrate peptide, provided in 96-well format. Optimal assay conditions have been identified for each substrate peptide using a phosphotyrosine-specific antibody to achieve signal-to noise ratios over 25.

Pathways & Proteins

Explore pathways + proteins related to this product.

For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
DELFIA is a registered trademark of PerkinElmer, Inc.