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7719
HTScan® EphB4 Kinase Assay Kit
Cellular Assay Kits
Assay Kit

HTScan® EphB4 Kinase Assay Kit #7719

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HTScan® EphB4 Kinase Assay Kit: Image 1
Western blot analysis of extracts from Jurkat cells treated with 1 mM pervanadate for 30 minutes prior to lysis, using P-Tyr-100 Phospho-Tyrosine Mouse mAb. Proteins were separated by 2-D electrophoresis prior to blotting.
HTScan® EphB4 Kinase Assay Kit: Image 2
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Phospho-Tyrosine Mouse mAb (P-Tyr-100).
HTScan® EphB4 Kinase Assay Kit: Image 3
Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Phospho-Tyrosine Mouse mAb (P-Tyr-100).
HTScan® EphB4 Kinase Assay Kit: Image 4
Immunohistochemical analysis of paraffin-embedded NCI-H1650 xenograft untreated (left) or lambda-phosphatase-treated (right), using Phospho-Tyrosine Mouse mAb (P-Tyr-100).
HTScan® EphB4 Kinase Assay Kit: Image 5
Immunohistochemical analysis of paraffin-embedded human B-cell non-Hodgkin lymphoma, untreated (left) or lambda phosphatase treated (right), using Phospho-Tyrosine Mouse mAb (P-Tyr-100).
HTScan® EphB4 Kinase Assay Kit: Image 6
Immunohistochemical analysis of paraffin-embedded human squamous cell lung carcinoma using Phospho-Tyrosine Mouse mAb (P-Tyr-100).
HTScan® EphB4 Kinase Assay Kit: Image 7
Immunohistochemical analysis using Phospho-Tyrosine Mouse mAb (P-Tyr-100) on SignalSlide™ Phospho-EGF Receptor IHC Controls #8102 (paraffin-embedded KYSE450 cell pellets, untreated (left) or EGF-treated (right)).
HTScan® EphB4 Kinase Assay Kit: Image 8
Immunohistochemical analysis of paraffin-embedded human soft tissue squamous cell carcinoma using Phospho-Tyrosine Mouse mAb (P-Tyr-100).
HTScan® EphB4 Kinase Assay Kit: Image 9
Immunohistochemical analysis of paraffin-embedded normal human kidney using Phospho-Tyrosine Mouse mAb (P-Tyr-100).
HTScan® EphB4 Kinase Assay Kit: Image 10
Immunohistochemical analysis of paraffin-embedded normal human lung using Phospho-Tyrosine Mouse mAb (P-Tyr-100).
HTScan® EphB4 Kinase Assay Kit: Image 11
Immunohistochemical analysis of paraffin-embedded NCI-H1650 xenograft using Phospho-Tyrosine Mouse mAb (P-Tyr-100).
HTScan® EphB4 Kinase Assay Kit: Image 12
Confocal immunofluorescent analysis of paraffin-embedded human lung adenocarcinoma using Phospho-Tyrosine Mouse mAb (P-Tyr-100) (red). Blue pseudocolor = DRAQ5® (fluorescent DNA dye).
HTScan® EphB4 Kinase Assay Kit: Image 13
Immunofluorescent analysis of Swiss NIH/3T3 cells, serum-starved and stimulated with lysophosphatidic acid (LPA) (10 µM for 10 minutes) and fixed with PFA, using Phospho-Tyrosine Mouse mAb (P-Tyr-100) (red) and phalloidin for F-actin (green) LPA causes heavy tyrosine phosphorylation of proteins in focal adhesions, present at the tips of actin stress fibers. (Provided by Dr. Harry Mellor, University of Bristol.)
HTScan® EphB4 Kinase Assay Kit: Image 14
Confocal immunofluorescence analysis of HeLa cells either stimulated with 20% serum (left) or untreated (right), using Phospho-Tyrosine Mouse mAb (P-Tyr-100) (red). Actin filaments have been labeled with fluorescein phalloidin. Blue pseudocolor = DRAQ5® (fluorescent DNA dye).
HTScan® EphB4 Kinase Assay Kit: Image 15
Flow cytometric analysis of K562 cells, untreated (green) or Gleevec®- treated (blue), using Phospho-Tyrosine Mouse mAb (P-Tyr-100) compared to a nonspecific negative control antibody (red).
Inquiry Info.# 7719

Product Description

The kit provides a means of performing kinase activity assays with recombinant human EphB4 kinase. It includes active EphB4 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.
MW (kDa) Peptide substrate, Biotin-FGFR-3 (Tyr724): 1,789 Daltons. GST-EphB4 Kinase: 77
Molecular Formula Peptide substrate, Biotin-FGFR-3 (Tyr724): 1,789 Daltons. GST-EphB4 Kinase: 77 kDa.

Background

The Eph receptors are the largest known family of receptor tyrosine kinases (RTKs). They can be divided into two groups based on sequence similarity and on their preference for a subset of ligands: EphA receptors bind to a glycosylphosphatidylinositol-anchored ephrin A ligand; EphB receptors bind to ephrin B proteins that have a transmembrane and cytoplasmic domain (1,2). Research studies have shown that Eph receptors and ligands may be involved in many diseases including cancer (3). Both ephrin A and B ligands have dual functions. As RTK ligands, ephrins stimulate the kinase activity of Eph receptors and activate signaling pathways in receptor-expressing cells. The ephrin extracellular domain is sufficient for this function as long as it is clustered (4). The second function of ephrins has been described as "reverse signaling", whereby the cytoplasmic domain becomes tyrosine phosphorylated, allowing interactions with other proteins that may activate signaling pathways in the ligand-expressing cells (5). Various stimuli can induce tyrosine phosphorylation of ephrin B, including binding to EphB receptors, activation of Src kinase, and stimulation by PDGF and FGF (6). Tyr324 and Tyr327 have been identified as major phosphorylation sites of ephrin B1 in vivo (7).
  1. Wilkinson, D.G. (2000) Int Rev Cytol 196, 177-244.
  2. Klein, R. (2001) Curr Opin Cell Biol 13, 196-203.
  3. Dodelet, V.C. and Pasquale, E.B. (2000) Oncogene 19, 5614-9.
  4. Holder, N. and Klein, R. (1999) Development 126, 2033-44.
  5. Brückner, K. et al. (1997) Science 275, 1640-3.
  6. Palmer, A. et al. (2002) Mol Cell 9, 725-37.
  7. Kalo, M.S. et al. (2001) J Biol Chem 276, 38940-8.

Pathways & Proteins

Explore pathways + proteins related to this product.

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