Human EphA3 HEK293 Overexpression Lysate

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Human EphA3 HEK293 Overexpression Lysate: Product Information

Product Description
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.
Expression Host
HEK293 Cells
Species
Human
Sequence Information
A DNA sequence encoding the human EPHA3 (Met1-Gln541) was expressed with a polyhistidine tag at the C-terminus.
Molecule Mass
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

Preparation Method
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.
Lysis Buffer
Modified RIPA Lysis Buffer: 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF.
Recommend Usage
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.
Sample Buffer
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.
Application
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.
Full Name
EPH receptor A3
References
  • Holder N, et al. (1999) Eph receptors and ephrins: effectors of morphogenesis. Development. 126(10):2033-44.
  • Wilkinson DG. (2000) Eph receptors and ephrins: regulators of guidance and assembly. Int Rev Cytol. 196:177-244.
  • Xu Q, et al. (2001) Roles of Eph receptors and ephrins in segmental patterning. Philos Trans R Soc Lond B Biol Sci. 355(1399):993-1002.
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