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Encompassing Amino Acids: 25-645
Application: Useful for binding assays and biotin-avidin pulldown assays.
Assay Conditions: A white 96-well plate was precoated with Anti-EGFR, Avi-Tag Antibody (#102551) (100 ng/well ). Blocking Buffer 3 (#79743) and PBST were used for blocking and washing steps, respectively. For protein titration a serial dilution of EGFR, His-Avi-Tag, Biotin-Labeled Recombinant (#102676) at the highest concentration of 4 ng/µl (200 ng/reaction) was prepared in 1x PP-02 Assay Buffer and added to the wells at Room Temperature (RT) for 1 hour. The plate was then washed again and incubated with Streptavidin-HRP diluted 1:1000 in Blocking Buffer 3 for 1 hour at RT. After the final wash, ELISA ECL substrate mix (#79670) was added to each well and chemiluminescence was read.
Background: EGFR (epidermal growth factor receptor) is the receptor for members of the EGF family and is a transmembrane glycoprotein that has tyrosine kinase activity. Binding of epidermal growth factor to EGFR induces receptor dimerization and tyrosine autophosphorylation and leads to cell proliferation, differentiation, motility, and cell survival. Activation of EGFR triggers mitogenic signaling in gastrointestinal mucosa, and its expression is upregulated in colon cancers and most neoplasms. EGFR activates the ERK (extracellular signal-regulated kinase)-signaling pathway in normal gastric epithelial and colon cancer cell lines. Inactivation of EGFR with selective inhibitors significantly reduces ERK2 activation, c-fos mRNA expression and cell proliferation, opening new therapeutic opportunities.
Description: Recombinant human EGFR (epidermal growth factor receptor), transcript variant 1, encompassing amino acids 25-645 of the extracellular domain. This construct contains a C-terminal His-tag (10xHis) followed by an Avi-Tag™. The recombinant protein was affinity purified. The recombinant protein was enzymatically biotinylated using the Avi-Tag™.
Format: Aqueous buffer solution
Formulation: 8 mM phosphate, pH 7.4, 110 mM NaCl, 2.2 mM KCl, and 20% glycerol
Genbank: NM_005228
Host Cell Line: HEK293
Purity: ≥85%
Storage Stability: At least 6 months at –80°C.
Tags: His-Avi
Target: EGFR
Uniprot: P00533
Unit Definition: Protein
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
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