HTScan® EGFR Kinase Assay Kit #7410
Inquiry Info. # 7410
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Product Information
Specificity / Sensitivity
Purified EGF-R kinase was quality controlled for purity using silver stain SDS-PAGE and Western blot. EGF-R kinase Vmax and Km values were measured to determine specific enzymatic activity (fig.5).
Assay conditions (time course [fig.1], kinase dose-dependence [fig.3], substrate dose-dependence [fig.4] and inhibitor sensitivity [fig.6]) for EGFR kinase activity were verified using the EGF-R substrate peptide provided in this kit.
Background
Description: The kit provides a means of performing enzymatic assays with active human EGF-R kinase. It includes active EGF-R kinase (supplied as a GST fusion protein), a biotinylated substrate peptide and a phospho-tyrosine monoclonal antibody for detection of the phosphorylated form of the substrate peptide.
Unit Definition: 10 Units is defined as the amount of EGF-R kinase required to maximally phosphorylate 75 pmol of PTP1B (Tyr66) #C03-1727 biotinylated substrate peptide in 30 minutes at 25ºC in a total reaction volume of 50 µl quantified by DELFIA®.
Peptide Core Sequence: NDY*IN
Molecular Weights: Peptide Substrate, Biotin-PTP1B (Tyr66): 2141 Daltons, GST-EGF-R Kinase domain: 91,163 Daltons
Storage: Antibodies are supplied in in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Do not aliquot the antibodies. Peptides are supplied at 6 µM in 0.001% DMSO. Enzymes are supplied in 50 mM Tris-HCL (pH 8.0), 100 mM NaCl, 5 mM DTT, 15 mM reduced glutathion and 20% glycerol. Store at -80ºC.
Keep enzymes on ice during use.
Avoid repeated freeze-thaw cycles.
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