Eck (EphA2, Mpk-5, Native Epithelial Cell Kinase, Eph receptor tyrosine kinase A2)

Eck (EphA2) is a member of the Eph receptor tyrosine kinase family which binds Ephrins A1-A5. A and B class Eph proteins have a common structural organization. The human EphA2 cDNA encodes a 976aa precursor including a 24aa signal sequence, a 510aa extra- cellular domain (ECD), a 24aa transmembrane segment and a 418 aa cytoplasmic domain. The ECD contains an N-terminal globular domain, a cysteine-rich domain and two fibronectin type III domains. The cytoplasmic domain contains a juxtamembrane motif with two tyrosine residues, which are the major autophosphorylation sites, a kinase domain, and a sterile alpha motif (SAM). The ECD of human EphA2 shares 90-94% aa sequence identity with mouse, bovine, and canine EphA2 and ~45% aa sequence identity with human EphA1, 3, 4, 5, 7 and 8. EphA2 becomes autophosphorylated following ligand binding and then interacts with SH2 domain- containing PI3-kinase to activate MAPK pathways. Reverse signaling is also propagated through the Ephrin ligand. Transcription of EphA2 is dependent on the expression of E-Cadherin and can be induced by p53 family transcription factors. EphA2 is upregulated in breast, prostate and colon cancer vascular endothelium. Its ligand, EphrinA1, is expressed by the local tumor cells. In some cases, EphA2 and EphrinA1 are expressed on the same blood vessels. EphA2 signaling cooperates with VEGF receptor signaling in promoting endothelial cell migration. The gene encoding human EphA2 maps to a region on chromosome 1 which is frequently deleted in neuroectodermal tumors.\n\nApplications: \nSuitable for use in Flow Cytometry, Western Blot and Immunocytochemistry. Other applications not tested.\nRecommended Dilutions:\nFlow Cytometry: 2.5ug labels 10e6 A431 cells.\nWestern Blot: 1ug/ml stains recombinant human EphA2 under non-reducing conditions only.\nImmunocytochemistry: 8-25ug/ml detects Eck on immersion fixed A431 cells. \nOptimal dilutions to be determined by the researcher.\nStorage and Stability:\nLyophilized powder may be stored at -20 degrees C. Stable for 12 months at -20 degrees C. Reconstitute with sterile PBS. Aliquot to avoid repeated freezing and thawing. Store at -20 degrees C. Reconstituted product is stable for 12 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

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