This Human DDR2 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of DDR2 protein (Cat: 10209-H02H) from the overexpression lysate was verified.
A DNA sequence encoding the extracellular domain (Met 1-Arg 399) of human DDR2 precursor (NP_001014796.1) was expressed with the fused Fc region of human IgG1 at the C-terminus.
The recombinant human DDR2/Fc is a disulfide-linked homodimeric protein. The reduced monomer consists of 616 amino acids and has a calculated molecular mass of 69.4 kDa. Due to glycosylation, rhDDR2/Fc monomer migrates as an approximately 87 kDa protein in SDS-PAGE under reducing conditions.
Human DDR2 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 DDR2 HEK293 Overexpression Lysate: 别称
Human CD167 Overexpression Lysate; Human MIG20a Overexpression Lysate; Human NTRKR3 Overexpression Lysate; Human TKT Overexpression Lysate; Human TYRO10 Overexpression Lysate
Discoidin domain receptor 2 (DDR2) or CD167b (cluster of differentiation 167b) is a kind of protein tyrosine kinases associated with cell proliferation and tumor metastasis, and collagen, identified as a ligand for DDR2, up-regulates matrix metallloproteinase 1 (MMP-1) and MMP-2 expression in cellular matrix. DDR2/CD167b was found to recognise the triple-helical region of collagen X as well as the NC1 domain. Binding to the collagenous region was dependent on the triple-helical conformation. DDR2/CD167b autophosphorylation was induced by the collagen X triple-helical region but not the NC1 domain, indicating that the triple-helical region of collagen X contains a specific DDR2 binding site that is capable of receptor activation. DDR2/CD167b is induced during stellate cell activation and implicate the phosphorylated receptor as a mediator of MMP-2 release and growth stimulation in response to type I collagen. Moreover, type I collagen-dependent upregulation of DDR2/CD167b expression establishes a positive feedback loop in activated stellate cells, leading to further proliferation and enhanced invasive activity.