This Mouse EphB4 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of EphB4 protein (Cat: 50582-M08H) from the overexpression lysate was verified.
A DNA sequence encoding the mouse EPHB4 (P54761-1) extracellular domain (Met 1-Ala 539) was fused with a polyhistidine tag at the C-terminus.
The secreted recombinant mouse EPHB4 comprises 535 amino acids and has a predicted molecular mass of 59 kDa. As a result of glycosylation, the apparent molecular mass of rm EPHB4/His monomer is approximately 70 kDa in SDS-PAGE under reducing conditions.
Mouse EphB4 HEK293 Overexpression Lysate: 使用指南
Cell lysate was prepared by homogenization of the over-expressed cells in ice-cold modified RIPA Lysis Buffer with cocktail of protease inhibitors (Sigma). Cell debris was removed by centrifugation. Protein concentration was determined by Bradford assay (Bio-Rad protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
Ephrin type-B receptor 4 is a protein that in humans is encoded by the EPHB4 gene. It is a single-pass type I membrane protein belonging to the ephrin receptor subfamily of protein kinase superfamily. Members of the ephrin and Eph family are local mediators of cell function through largely contact-dependent processes in development and in maturity. Furthermore, EphB4 protein and the corresponding ligand Ephrin-B2 contribute to tumor growth in various human tumors. EphB4 protein has tumor suppressor activities and that regulation of cell proliferation, extracellular matrix remodeling, and invasive potential are important mechanisms of tumor suppression. Therefore, Ephrin-B2/EphB4 may be recognized as a novel prognostic indicator for cancers.
EPH receptor B4
Actin Dynamics Signaling Pathway
G Protein-coupled Receptors Signaling
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