Recombinant human EGFR /ErbB1 Protein, Fc Tag, 100µg

Background
The epidermal growth factor receptor (EGFR; ErbB-1; HER1 in humans) is the cell-surface receptor for members of the epidermal growth factor family (EGF-family) of extracellular protein ligands.[1] The epidermal growth factor receptor is a member of the ErbB family of receptors, a subfamily of four closely related receptor tyrosine kinases: EGFR (ErbB-1), HER2/c-neu (ErbB-2), Her 3 (ErbB-3) and Her 4 (ErbB-4). Mutations affecting EGFR expression or activity could result in cancer.[2]

Source
Recombinant Human EGFR /ErbB1 Protein, With C-Fc Tag (rhEGFR-Fc) Leu 25 - Ser 645 (Accession # NP_005219) was produced in human 293 cells (HEK293).

Molecular Characterization
rhEGFR-Fc, fused with Fc fragment of human IgG1 at the C-terminus, has a calculated MW of 95 kDa. The predicted N-terminus is Leu 25. DTT-reduced Protein migrates as 130-140 kDa due to glycosylation.

Endotoxin
Less than 1.0 EU per µg of the rhEGFR-Fc by the LAL method.

Purity
>95% as determined by SDS-PAGE.

Formulation
Lyophilized from 0.22 µm filtered solution in 50 mM tris, 100 mM glycine, pH7.0. Normally Mannitol or Trehalose are added as protectants before lyophilization.

Reconstitution
See Certificate of Analysis for details of reconstitution instruction and specific concentration.

Storage
Avoid repeated freeze-thaw cycles.
No activity loss was observed after storage at:
In lyophilized state for 1 year (4°C-8°C); After reconstitution under sterile conditions for 1 month (4°C-8°C) or 3 months (-20°C to -70°C).

Bioactivity
Measured by its binding ability in a functional ELISA. Immobilized Human EGF, Tag Free (Catalog # EGF-H4114) at 10μg/mL(100 μL/well) can bind Human EGF R, Fc Tag (Catalog # EGR-H5252) with a linear range of 0.2-3.2 μg/mL. Measured by its ability to bind Human EGF, Tag Free (Cat# EGF-H4114) in the SPR assay (Biacore 2000) with an estimated KD < 1nM.Validated CetuxiMab binding in a functional ELISA (routinely tested).

References

(1) Herbst RS, 2004, Int. J. Radiat. Oncol. Biol. Phys. 59 (2 Suppl): 21–6.
(2) Zhang H, et al.,2007, Journal of Clinical Investigation 117(8):2051-2058.