This Mouse EphA1 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of EphA1 protein (Cat: 50827-M08H) from the overexpression lysate was verified.
A DNA sequence encoding the extracellular domain of mouse EPHA1 (Q60750) (Met 1-Glu 548) was expressed, with a C-terminal polyhistidine tag.
The secreted recombinant mouse EPHA1 comprises 535 amino acids and has a calculated molecular mass of 58.6 kDa. As a result of glycosylation, the apparent molecular mass of rmEPHA1 is approximately 68 kDa in SDS-PAGE under reducing conditions.
Mouse EphA1 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.
EPHA1 or EPH receptor A1 belongs to the ephrin receptor subfamily of the protein-tyrosine kinase family. Receptors in the EPH subfamily typically have a single kinase domain and an extracellular region containing a Cys-rich domain and 2 fibronectin type III repeats. An important role of Eph receptors and their ligands ephrins is to mediate cell-contact-dependent repulsion. Eph receptors and ephrins also act at boundaries to channel neuronal growth cones along specific pathways, restrict the migration of neural crest cells, and via bidirectional signaling prevent intermingling between hindbrain segments. Eph receptors and ephrins can also trigger an adhesive response of endothelial cells and are required for the remodeling of blood vessels. Eph receptors and ephrins have emerged as key regulators of the repulsion and adhesion of cells that underlie the establishment, maintainence, and remodeling of patterns of cellular organization. The ephrins and Eph receptors are implicated as positional labels that may guide the development of neural topographic maps.
EPH receptor A1
Actin Dynamics Signaling Pathway
G Protein-coupled Receptors Signaling
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Zhou R. (1998) The Eph family receptors and ligands. Pharmacol. 77 (3): 151-81.