This Mouse DDR1 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of DDR1 protein (Cat: 50829-M02H) from the overexpression lysate was verified.
A DNA sequence encoding the extracellular domain of mouse DDR1 (NP_766550.1) (Met 1-Thr 414) was fused with the Fc region of human IgG1 at the C-terminus
The secreted recombinant mouse DDR1/Fc is a disulfide-linked homodimer. The reduced monomer comprises 636 amino acids and has a calculated molecular mass of 71 kDa. As a result of glycosylation, the apparent molecular mass of rmDDR1/Fc monomer is approximately 80-90 kDa in SDS-PAGE under reducing conditions.
Mouse DDR1 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.
Discoidin domain receptor family, member 1 (DDR1), also known as or CD167a (cluster of differentiation 167a), and Mammary carcinoma kinase 10 (MCK10), belongs to a subfamily of tyrosine kinase receptors with an extracellular domain homologous to Dictyostellium discoideum protein discoidin 1. Receptor tyrosine kinases play a key role in the communication of cells with their microenvironment. These kinases are involved in the regulation of cell growth, differentiation and metabolism. Expression of DDR1/MCK10/CD167 is restricted to epithelial cells, particularly in the kidney, lung, gastrointestinal tract, and brain. In addition, it has been shown to be significantly overexpressed in several human tumors. DDR1/MCK10/CD167 plays an important role in regulating attachment to collagen, chemotaxis, proliferation, and MMP production in smooth muscle cells. DDR1 functions in a feedforward loop to increase p53 levels and at least some of its effectors. Inhibition of DDR1 function resulted in strikingly increased apoptosis of wild-type p53-containing cells in response to genotoxic stress through a caspase-dependent pathway.