Mouse EGFR HEK293 Overexpression Lysate

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Mouse EGFR HEK293 Overexpression Lysate: 产品信息

产品描述
This Mouse EGFR overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of EGFR protein (Cat: 51091-M02H) from the overexpression lysate was verified.
表达宿主
HEK293 Cells
种属
Mouse
蛋白构建信息
A DNA sequence encoding the extracellular domain of mouse EGFR (Q01279) (Met 1-Ser 647) was fused with the Fc region of human IgG1 at the C-terminus.
分子量
The secreted recombinant mouse EGFR/Fc is a disulfide-linked homodimer. The reduced monomer comprises 864 amino acids and has a calculated molecular mass of 96 kDa. As a result of glycosylation, the apparent molecular mass of rmEGFR/Fc monomer is approximately 130 kDa in SDS-PAGE under reducing conditions.

Mouse EGFR HEK293 Overexpression Lysate: 使用指南

制备方法
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.
裂解缓冲液
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.
使用建议
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.
缓冲液
1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
稳定性 & 储存条件
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.
应用
Western Blot (WB)
Optimal dilutions/concentrations should be determined by the end user.

Mouse EGFR HEK293 Overexpression Lysate: 别称

Mouse 9030024J15Rik Overexpression Lysate; Mouse AI552599 Overexpression Lysate; Mouse Erbb Overexpression Lysate; Mouse Errb1 Overexpression Lysate; Mouse Errp Overexpression Lysate; Mouse wa-2 Overexpression Lysate; Mouse wa2 Overexpression Lysate; Mouse Wa5 Overexpression Lysate

EGFR 背景信息

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.
全称
epidermal growth factor receptor
参考文献
  • Schlessinger, J. (2000) Cell signaling by receptor tyrosine kinases. Cell 103(2): 211-25.
  • Giaccone, G. (2005) HER1/EGFR-targeted agents: predicting the future for patients with unpredictable outcomes to therapy. Ann. Oncol. 16(4): 538-48.
  • Yarden, Y., et al. (2001) Untangling the ErbB signalling network. Nat. Rev. Mol. Cell. Biol. 2(2): 127-37.
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