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Human CD136 / MST1R HEK293 Cell Lysate (WB positive control)

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Human MST1R Transfected / Overexpression Cell Lysate Product Information
Expressed Host:Human Cells
Product Description:Human Cell lysate that Human MST1R / CD136 transfected / overexpressed for Western blot (WB) positive control. The whole cell lysate is provided in 1X Sample Buffer (1X modified RIPA buffer+1X SDS loading buffer).
Sequence information:A DNA sequence encoding the amino acid sequence (Met 1-Leu 571) of human CD136 (Q04912) extracellular domain was fused with a polyhistidine tag at the C-terminus.
Predicted N Terminal:Glu 25 & Gly 310
Molecule Mass:The secreted recombinant human CD136 consists of 558 amino acids, including the α chain (Glu 25-Arg 309) and the polyhistidine-tagged β chain (Gly 310-Leu 517), and predictes a molecular mass of 60 kDa. (30+30 kDa). As a result of glycosylation, in SDS-PAGE under reducing conditions, the apparent molecular mass of rhCD136 is approximately 70 kDa and 37 kDa, corresponding to the single chain and the cleaved two subunits respectively.
Human MST1R Transfected / Overexpression Cell Lysate Usage Guide
Preparation Method: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.
Lysis Buffer: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.
Quality Control Testing:12.5% SDS-PAGE Stained with Coomassie Blue after protein purification.
Stability:Samples are stable for up to twelve months from date of receipt.
Recommend Usage: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℃.
Storage Buffer:1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
Storage Instruction:Store at 4℃. After re-dissolution, aliquot and store at -80℃.
Application notes:Western blot (WB): Use at an assay dependent dilution.
Other Applications: Not tested.
Optimal dilutions/concentrations should be determined by the end user.
CD136/MST1R Background

The tyrosine kinase receptor, macrophage-stimulating 1 receptor (MST1R), a c-met-related tyrosine kinase, also known as the Ron receptor or CD136, controls cell survival and motility programs related to invasive growth. As tyrosine kinase receptor comprised of an extra-cellular domain, MST1R protein contains the ligand binding pocket and an intracellular region where the kinase domain is located. MST1R signaling may be involved in the regulation of macrophage and T-lymphocyte activation in vivo during injury. This assessment of gene expression indicates the importance of genetic factors in contributing to lung injury, and points to strategies for intervention in the progression of inflammatory diseases. It had been shown that MST1R/CD136 plays a critical role in Ni-induced lung injury in mice. The overexpression of MSP, MT-SP1, and MST1R was a strong independent indicator of both metastasis and death in human breast cancer patients and significantly increased the accuracy of an existing gene expression signature for poor prognosis. Stimulation of MST1R leads to its transphosphorylation and the ultimate activation of numerous intracellular signaling pathways, such as the classical mitogen-activated protein kinase pathway, the phosphotidylinositol (PI)3-kinase pathway, and the JNK pathway.

Human CD136/MST1R References
  • Ronsin C, et al. (1993) A novel putative receptor protein tyrosine kinase of the met family. Oncogene. 8: 1195-1202.
  • McDowell SA, et al. (2002) The role of the receptor tyrosine kinase Ron in nickel-induced acute lung injury. Am J Respir Cell Mol Biol. 26(1): 99-104.
  • Angeloni D, et al. (2003) C to A single nucleotide polymorphism in intron 18 of the human MST1R (RON) gene that maps at 3p21.3. Mol Cell Probes. 17(2-3): 55-7.
  • Mallakin A, et al. (2006) Gene expression profiles of Mst1r-deficient mice during nickel-induced acute lung injury. Am J Respir Cell Mol Biol. 34(1): 15-27.
  • Welm AL, et al. (2007) The macrophage-stimulating protein pathway promotes metastasis in a mouse model for breast cancer and predicts poor prognosis in humans. Proc Natl Acad Sci U S A. 104(18): 7570-5.
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    Catalog: 11608-H08HL-300
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