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7725
HTScan® HER4/ErbB4 Kinase Assay Kit

HTScan® HER4/ErbB4 Kinase Assay Kit #7725

This product is discontinued

Kinase Assay - Radiometric Image 1

Figure 1. HER4/ErbB4 kinase activity was measured in a radiometric assay using the following reaction conditions: 60 mM HEPES-NaOH, pH 7.5, 3 mM MgCl2, 3 mM MnCl2, 3 µM Na-orthovanadate, 1.2 mM DTT, ATP (variable), 2.5 µg/50 µl PEG20.000, Substrate: PolyAEKY, 5 µg/50 µl, recombinant HER4/ErbB4: 50 ng/50 µl.

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Kinase Assay - DELFIA Image 2

Figure 3. Dose dependence curve of HER4/ErbB4 kinase activity: DELFIA® data generated using Phospho-Tyrosine mAb (P-Tyr-100) #9411 to detect phosphorylation of substrate peptide (#1315) by HER4/ErbB4 kinase. In a 50 µl reaction, increasing amounts of HER4/ErbB4 and 1.5 µM substrate peptide were used per reaction at room temperature for 30 minutes. (DELFIA® is a registered trademark of PerkinElmer, Inc.)

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Kinase Assay - DELFIA Image 3

Figure 5. Staurosporine inhibition of HER4/ErbB4 kinase activity: DELFIA® data generated using Phospho-Tyrosine mAb (P-Tyr-100) #9411 to detect phosphorylation of HER4/ErbB4 substrate peptide (#1315) by HER4/ErbB4 kinase. In a 50 µl reaction, 50 ng HER4/ErbB4, 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.)

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Kinase Assay - DELFIA Image 4

Figure 4. Peptide concentration dependence of HER4/ErbB4 kinase activity: DELFIA® data generated using Phospho-Tyrosine mAb (P-Tyr-100) #9411 to detect phosphorylation of substrate peptide (#1315) by HER4/ErbB4 kinase. In a 50 µl reaction, 50 ng of HER4/ErbB4 and increasing concentrations of substrate peptide were used per reaction at room temperature for 30 minutes. (DELFIA® is a registered trademark of PerkinElmer, Inc.)

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Kinase Assay - DELFIA Image 5

Figure 2. Time course of HER4/ErbB4 kinase activity: DELFIA® data generated using Phospho-Tyrosine mAb (P-Tyr-100) #9411 to detect phosphorylation of HER4/ErbB4 substrate peptide (#1315) by HER4/ErbB4 kinase. In a 50 µl reaction, 50 ng HER4/ErbB4 and 1.5 µM substrate peptide were used per reaction. (DELFIA® is a registered trademark of PerkinElmer, Inc.)

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Product Includes Quantity (with Count) Solution Color
DTT (1000X) 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 HER4/ErbB4 kinase. It includes active HER4/ErbB4 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-peptide: 2,166 Daltons. GST-HER4/ErbB4 Kinase: 102 kDa.

Peptide Core Sequence:

DIY*AE

Research studies have implicated the HER/ErbB receptor tyrosine kinase family in normal development, cardiac function and cancer (1,2). HER4/ErbB4, like other family members, has four ectodomains, a single transmembrane domain and a cytoplasmic tail containing the active tyrosine kinase domain (3). By binding to neuregulins and/or EGF family ligands, ErbB4 forms either a homodimer or heterodimer with other ErbB family members, which results in receptor activation and signaling (3). ErbB4 is ubiquitously expressed with the highest expression occurring in brain and heart. The expression of ErbB4 in breast cancer, pediatric brain cancer and other types of carcinomas has been reported in research studies suggesting that ErbB4 expression is involved in both normal tissue development and carcinogenesis (3).

  1. Holbro, T. and Hynes, N.E. (2004) Annu Rev Pharmacol Toxicol 44, 195-217.
  2. Roskoski, R. (2004) Biochem Biophys Res Commun 319, 1-11.
  3. Carpenter, G. (2003) Exp Cell Res 284, 66-77.
Entrez-Gene Id
2066
Swiss-Prot Acc.
Q15303
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.

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