Human Her2 / ERBB2 HEK293 Overexpression Lysate

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Human Her2 / ERBB2 HEK293 Overexpression Lysate: 产品信息

产品描述
This Human Her2 / ERBB2 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of Her2 / ERBB2 protein (Cat: 10004-H02H) from the overexpression lysate was verified.
表达宿主
HEK293 Cells
种属
Human
蛋白构建信息
A DNA sequence encoding the extracellular domain (Met 1-Thr 652) of human ErbB2 (NP_004439.2) was fused with the Fc region of human IgG1 at the C-terminus.
分子量
The recombinant human ErbB2/Fc chimera is a disulfide-linked homodimeric protein generated by proteolytic removal of the signal peptide. The monomer comprises 868 amino acids and has a calculated molecular mass of 96.1 kDa. As a result of glycosylation, the monomer migrates as an approximately 130-140 kDa protein in SDS-PAGE under reducing conditions.

Human Her2 / ERBB2 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.

Human Her2 / ERBB2 HEK293 Overexpression Lysate: 别称

Human CD340 Overexpression Lysate; Human ENV Overexpression Lysate; Human ENVW Overexpression Lysate; Human ERVWE1 Overexpression Lysate; Human HER-2 Overexpression Lysate; Human HER-2/neu Overexpression Lysate; Human HER2 Overexpression Lysate; Human HERV-7q Overexpression Lysate; Human HERV-W-ENV Overexpression Lysate; Human HERV7Q Overexpression Lysate; Human HERVW Overexpression Lysate; Human HERVWENV Overexpression Lysate; Human MLN 19 Overexpression Lysate; Human MLN19 Overexpression Lysate; Human NEU Overexpression Lysate; Human NGL Overexpression Lysate; Human TKR1 Overexpression Lysate

Her2 / ERBB2 背景信息

Epidermal growth factor receptor 2 (HER2), also known as ErbB2, NEU, and CD34, is a type I membrane glycoprotein, and belongs to the epidermal growth factor (EGF) receptor family. HER2 protein cannot bind growth factors due to the lacking of ligand binding domain of its own and autoinhibited constitutively. However, HER2 forms a heterodimer with other ligand-bound EGF receptor family members, therefore stabilizes ligand binding and enhances kinase-mediated activation of downstream molecules. HER2 plays a key role in development, cell proliferation and differentiation. HER2 gene has been reported to associate with malignancy and a poor prognosis in numerous carcinomas, including breast, prostate, ovarian, lung cancers and so on.
全称
erb-b2 receptor tyrosine kinase 2
参考文献
  • Krawczyk N, et al. (2009) HER2 status on persistent disseminated tumor cells after adjuvant therapy may differ from initial HER2 status on primary tumor. Anticancer Res. 29(10): 4019-24.
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