Buy 3 Get a 4th Free* | Learn More >>
7058
HTScan® HER2/ErbB2 Kinase Assay Kit

HTScan® HER2/ErbB2 Kinase Assay Kit #7058

This product is discontinued

Kinase Assay - Radiometric Image 1

Figure 1. HER2/ErbB2 kinase activity was measured in a radiometric assay using the following reaction conditions: 5 mM MOPS, pH 7.2, 2.5 mM β-glycerophosphate, 5 mM MnCl2, 1 mM EGTA, 0.4 mM EDTA, 4 mM MgCl2, 0.05 mM DTT, 50 μM ATP, Substrate: Poly(Glu-Tyr), 400 ng/μL, and recombinant HER2/ErbB2: variable.

Learn more about how we get our images
Kinase Assay - DELFIA Image 2

Figure 3. Dose dependence curve of HER2/ErbB2 kinase activity: DELFIA® data generated using Phospho-Tyrosine mAb (P-Tyr-100) #9411 to detect phosphorylation of substrate peptide (#1305) by HER2/ErbB2 kinase. In a 50 µl reaction, increasing amounts of HER2/ErbB2 and 1.5 µM substrate peptide were used per reaction at room temperature for 30 minutes. (DELFIA® is a registered trademark of PerkinElmer, Inc.)

Learn more about how we get our images
Kinase Assay - DELFIA Image 3

Figure 5. Staurosporine inhibition of HER2/ErbB2 kinase activity: DELFIA® data generated using Phospho-Tyrosine mAb (P-Tyr-100) #9411 to detect phosphorylation of HER2/ErbB2 substrate peptide (#1305) by HER2/ErbB2 kinase. In a 50 µl reaction, 100 ng HER2/ErbB2, 1.5 µM substrate peptide, 20 µM ATP and increasing amounts of staurosporine were used per reaction at room temperature for 30 minutes. (DELFIA® is a registered trademark of PerkinElmer, Inc.)

Learn more about how we get our images
Kinase Assay - DELFIA Image 4

Figure 4. Peptide concentration dependence of HER2/ErbB2 kinase activity: DELFIA® data generated using Phospho-Tyrosine mAb (P-Tyr-100) #9411 to detect phosphorylation of substrate peptide (#1305) by HER2/ErbB2 kinase. In a 50 µl reaction, 50 ng of HER2/ErbB2 and increasing concentrations of substrate peptide were used per reaction at room temperature for 30 minutes. (DELFIA® is a registered trademark of PerkinElmer, Inc.)

Learn more about how we get our images
Kinase Assay - DELFIA Image 5

Figure 2. Time course of HER2/ErbB2 kinase activity: DELFIA® data generated using Phospho-Tyrosine mAb (P-Tyr-100) #9411 to detect phosphorylation of HER2/ErbB2 substrate peptide (#1305) by HER2/ErbB2 kinase. In a 50 µl reaction, 50 ng HER2/ErbB2 and 1.5 µM substrate peptide were used per reaction. (DELFIA® is a registered trademark of PerkinElmer, Inc.)

Learn more about how we get our images
Product Includes Quantity (with Count) Solution Color
DTT (1000X, 1.25 M) 1 x 80 µl
ATP (10 mM) 9804 1 x 1 ml

The kit provides a means of performing kinase activity assays with recombinant human HER2/ErbB2 kinase. It includes active HER2/ErbB2 kinase (supplied as a GST fusion protein), a biotinylated peptide substrate and a phospho-tyrosine antibody for detection of the phosphorylated form of the substrate peptide.

Molecular Formula:

Peptide substrate, Biotin-FLT3 (Tyr589) peptide: 1,945 Daltons. GST-HER2/ErbB2 Kinase: 116 kDa.

Peptide Core Sequence:

NEY*FY*V

The ErbB2 (HER2) proto-oncogene encodes a 185 kDa transmembrane, receptor-like glycoprotein with intrinsic tyrosine kinase activity (1). While ErbB2 lacks an identified ligand, ErbB2 kinase activity can be activated in the absence of a ligand when overexpressed and through heteromeric associations with other ErbB family members (2). Amplification of the ErbB2 gene and overexpression of its product are detected in almost 40% of human breast cancers (3). Binding of the c-Cbl ubiquitin ligase to ErbB2 at Tyr1112 leads to ErbB2 poly-ubiquitination and enhances degradation of this kinase (4). ErbB2 is a key therapeutic target in the treatment of breast cancer and other carcinomas and targeting the regulation of ErbB2 degradation by the c-Cbl-regulated proteolytic pathway is one potential therapeutic strategy. Phosphorylation of the kinase domain residue Tyr877 of ErbB2 (homologous to Tyr416 of pp60c-Src) may be involved in regulating ErbB2 biological activity. The major autophosphorylation sites in ErbB2 are Tyr1248 and Tyr1221/1222; phosphorylation of these sites couples ErbB2 to the Ras-Raf-MAP kinase signal transduction pathway (1,5).

  1. Muthuswamy, S.K. et al. (1999) Mol Cell Biol 19, 6845-57.
  2. Qian, X. et al. (1994) Proc Natl Acad Sci USA 91, 1500-4.
  3. Dittadi, R. and Gion, M. (2000) J Natl Cancer Inst 92, 1443-4.
  4. Klapper, L.N. et al. (2000) Cancer Res 60, 3384-8.
  5. Kwon, Y.K. et al. (1997) J Neurosci 17, 8293-9.
Entrez-Gene Id
2064
Swiss-Prot Acc.
P04626
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.

Upstream / Downstream

pathwayImage

Explore pathways related to this product.