This Mouse Ephrin B3 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of Ephrin B3 protein (Cat: 51147-M08H) from the overexpression lysate was verified.
A DNA sequence encoding the mouse Efnb3 (NP_031937.1) (Met1-Ala227) was expressed with a polyhistidine tag at the C-terminus.
The recombinant mouse Efnb3 consists 211 amino acids and predicts a molecular mass of 23.5 kDa.
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 B3 belongs to the ephrin family. Ephrins are divided into the ephrin-A (EFNA) class, which are anchored to the membrane by a glycosylphosphatidylinositol linkage, and the ephrin-B (EFNB) class, which are transmembrane proteins. Ephrin B3 is important in brain development as well as in its maintenance. It is especially important for forebrain function since its expression levels were particularly high in several forebrain subregions compared to other brain subregions. Ephrin B3 binds to, and induce the collapse of, commissural axons/growth cones in vitro. It may play a role in constraining the orientation of longitudinally projecting axons.
Takemoto M, et al. (2002) Ephrin-B3-EphA4 interactions regulate the growth of specific thalamocortical axon populations in vitro. Eur J Neurosci. 16(6):1168-72.
Brckner K, et al. (1999) EphrinB ligands recruit GRIP family PDZ adaptor proteins into raft membrane microdomains. Neuron. 22(3):511-24.
Bergemann A, et al. (1998) Ephrin-B3, a ligand for the receptor EphB3, expressed at the midline of the developing neural tube. Oncogene. 16(4):471-80.