Human EGFR Baculovirus-Insect Overexpression Lysate: Product Information
This Human EGFR overexpression lysate was created in Baculovirus-Insect Cells and intented for use as a Western blot (WB) positive control. Purification of EGFR protein (Cat: 29662-H08B) from the overexpression lysate was verified.
A DNA sequence encoding the human EGFRvIII (NP_001333870.1) (Met1-Lys375) was expressed with a polyhistidine tag at the C-terminus.
The recombinant human EGFRvIII consists of 359 amino acids and predicts a molecular mass of 39.3 kDa.
Human EGFR Baculovirus-Insect Overexpression Lysate: Usage Guide
Cell lysate was prepared by homogenization 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.
3. Store the lyophilized cell lysate at 4℃. After re-dissolution, recommend to aliquot it into smaller quantities and store at -80℃.
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 EGFR Baculovirus-Insect Overexpression Lysate: Synonyms
Human ERBB Overexpression Lysate; Human ERBB1 Overexpression Lysate; Human HER1 Overexpression Lysate; Human mENA Overexpression Lysate; Human NISBD2 Overexpression Lysate; Human PIG61 Overexpression Lysate
EGFR Background Information
As a member of the epidermal growth factor receptor (EGFR) family, EGFR protein is type I transmembrane glycoprotein that binds a subset of EGF family ligands including EGF, amphiregulin, TGF-α, betacellulin, etc. EGFR protein plays a crucial role in signaling pathway in the regulation of cell proliferation, survival and differentiation. Binding of a ligand induces EGFR protein homo- or heterodimerization, the subsequent tyrosine autophosphorylation and initiates various down stream pathways (MAPK, PI3K/PKB and STAT). In addition, EGFR signaling also has been shown to exert action on carcinogenesis and disease progression, and thus EGFR protein is proposed as a target for cancer therapy currently.
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