Human EPOR & CD131 Heterodimer HEK293 Overexpression Lysate

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Human EPOR & CD131 Heterodimer HEK293 Overexpression Lysate: Product Information

Product Description
This Human EPOR & CD131 Heterodimer overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of EPOR & CD131 Heterodimer protein (Cat: CT010-H02H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Species
Human
Sequence Information
A DNA sequence encoding the extracellular domain (Met 1-Pro 250) of human EPOR (NP_000112.1) linked with the extracellular domain (Trp 17-Trp 443) of human CD131 (NP_000386.1) by a peptide linker was fused with the Fc region of human IgG1 at the C-terminus.
Molecule Mass
The recombinant human EPOR&CD131/Fc chimera is a disulfide-linked homodimer of EPOR&CD131-Fc heterodimer. The reduced EPOR& CD131-Fc comprises 906 amino acids and has a calculated molecular mass of 101 kDa. As a result of glycosylation, the apparent molecular mass of the recombinant protein is approximately 125 kDa in SDS-PAGE under reducing conditions.

Human EPOR & CD131 Heterodimer 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 EPOR & CD131 Heterodimer HEK293 Overexpression Lysate: Alternative Names

Human EPO-R Overexpression Lysate

EPOR & CD131 Heterodimer Background Information

Full Name
erythropoietin receptor
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