Human c-MET Baculovirus-Insect cells Overexpression Lysate: Product Information
This Human c-MET overexpression lysate was created in Baculovirus-Insect cells and intented for use as a Western blot (WB) positive control. Purification of c-MET protein (Cat: 10692-H20B1) from the overexpression lysate was verified.
A DNA sequence encoding the human MET (P08581-1) (Lys956-Ser1390) was fused with the N-terminal polyhistidine-tagged GST tag at the N-terminus.
The recombinant human MET /GST chimera consists of 672 amino acids and has a calculated molecular mass of 76.8 kDa. The recombinant protein migrates as an approximately 68 kDa band in SDS-PAGE under reducing conditions.
Human c-MET Baculovirus-Insect cells Overexpression Lysate: Usage Guide
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).
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 c-MET Baculovirus-Insect cells Overexpression Lysate: Alternative Names
Human AUTS9 Overexpression Lysate; Human c-Met Overexpression Lysate; Human DFNB97 Overexpression Lysate; Human HGFR Overexpression Lysate; Human RCCP2 Overexpression Lysate
c-MET Background Information
Hepatocyte growth factor receptor (HGFR), also known as c-Met or mesenchymal-epithelial transition factor (MET), is a receptor tyrosine kinase (RTK) that has been shown to be 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.
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