Human EphA3 HEK293 Overexpression Lysate: Product Information
This Human EphA3 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of EphA3 protein (Cat: 11459-H08H) from the overexpression lysate was verified.
A DNA sequence encoding the human EPHA3 (Met1-Gln541) was expressed with a polyhistidine tag at the C-terminus.
The recombinant human EPHA3 consists of 532 amino acids and predicts a molecular mass of 60.3 KDa. It migrates as an approximately 65-70 KDa band in SDS-PAGE under reducing conditions.
Human EphA3 HEK293 Overexpression Lysate: Usage Guide
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.
1. Centrifuge the tube for a few seconds and ensure the pellet at the bottom of the tube.
2. Re-dissolve the pellet using 200μL pure water and boil for 2-5 min.
1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
Stability & Storage
Store at 4℃ for up to twelve months from date of receipt. After re-dissolution, aliquot and store at -80℃ for up to twelve months. Avoid repeated freeze-thaw cycles.
Western Blot (WB) Optimal dilutions/concentrations should be determined by the end user.
Human EphA3 HEK293 Overexpression Lysate: Alternative Names
Human EK4 Overexpression Lysate; Human ETK Overexpression Lysate; Human ETK1 Overexpression Lysate; Human HEK Overexpression Lysate; Human HEK4 Overexpression Lysate; Human TYRO4 Overexpression Lysate
EphA3 Background Information
EPHA3 gene belongs to the ephrin receptor subfamily of the protein-tyrosine kinase family. EPH and EPH-related receptors have been implicated in mediating developmental events, particularly in the nervous system. The ephrin receptors are divided into 2 groups based on the similarity of their extracellular domain sequences and their affinities for binding ephrin-A and ephrin-B ligands. EPHA3 gene encodes a protein that binds ephrin-A ligands. EPHA3 is involved in the retinotectal mapping of neurons. It may also control the segregation but not the guidance of motor and sensory axons during neuromuscular circuit development.