This Human c-MET overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of c-MET protein (Cat: 10692-H08H) from the overexpression lysate was verified.
A DNA sequence encoding the extracellular domain of human c-Met (NP_000236.2) (Met1-Thr932) was fused with a polyhistidine tag at the C-terminus.
The mature form of recombinant human c-Met is a disulfide-linked heterodimer composed of proteolytically cleaved α and β subunits. Each α and β subunit together consists of 919 amino acids and has a predicted molecular mass of 103 (α=33 +β=70) kDa. As a result of glycosylation, rh c-MET heterodimer thus migrates with apparent molecular mass of approximately 45 kDa and 85 kDa respectively in SDS-PAGE under reducing conditions.
Human c-MET HEK293 Overexpression Lysate: 使用指南
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.
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.
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.
Western Blot (WB) Optimal dilutions/concentrations should be determined by the end user.
Human c-MET HEK293 Overexpression Lysate: 别称
Human AUTS9 Overexpression Lysate; Human c-Met Overexpression Lysate; Human DFNB97 Overexpression Lysate; Human HGFR Overexpression Lysate; Human RCCP2 Overexpression Lysate
Hepatocyte growth factor receptor (HGFR), also known as c-Met or mesenchymal-epithelial transition factor (MET), is a receptor tyrosine kinase (RTK) that is overexpressed and/or mutated in a variety of malignancies. HGFR protein is produced as a single-chain precursor, and HGF is the only known ligand. Normal HGF/HGFR signaling is essential for embryonic development, tissue repair, or wound healing, whereas aberrantly active HGFR has been strongly implicated in tumorigenesis, particularly in the development of invasive and metastatic phenotypes. HGFR protein is a multifaceted regulator of growth, motility, and invasion, and is normally expressed by cells of epithelial origin. Preclinical studies suggest that targeting aberrant HGFR signaling could be an attractive therapy in cancer.