This Mouse MDGA2 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of MDGA2 protein (Cat: 50431-M08H) from the overexpression lysate was verified.
A DNA sequence encoding the mouse MDGA2 isoform 1 (P60755-1) without the propeptide (Met 1-Asp 924) was expressed, fused with a polyhistidine tag at the C-terminus.
The secreted recombinant mouse MDGA2 comprises 910 amino acids and has a predicted molecular mass of 103 kDa. As a result of glycosylation, the apparent molecular mass of rm MDGA2 is approximately 110-120 kDa in SDS-PAGE under reducing conditions.
Mouse MDGA2 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.
MAM domain-containing glycosylphosphatidylinositol anchor protein 2, also known as MAM domain-containing protein 1, MDGA2 and MAMDC1, is a cell membrane protein which contains sixIg-like (immunoglobulin-like) domains and oneMAM domain. Analyses of the full-length coding region of MDGA1 and MDGA2 indicate that they encode proteins that comprise a novel subgroup of the Ig superfamily and have a unique structural organization consisting of six immunoglobulin (Ig)-like domains followed by a single MAM domain. Biochemical characterization demonstrates that MDGA1 and MDGA2 proteins are highly glycosylated, and that MDGA1 is tethered to the cell membrane by a GPI anchor. The MDGAs are differentially expressed by subpopulations of neurons in both the central and peripheral nervous systems, including neurons of the basilar pons, inferior olive, cerebellum, cerebral cortex, olfactory bulb, spinal cord, and dorsal root and trigeminal ganglia. The similarity of MDGAs to other Ig-containing molecules and their temporal-spatial patterns of expression within restricted neuronal populations, for example migrating pontine neurons and D1 spinal interneurons, suggest a role for these novel proteins in regulating neuronal migration, as well as other aspects of neural development, including axon guidance.
MAM domain containing glycosylphosphatidylinositol anchor 2