This Mouse Neuroserpin overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of Neuroserpin protein (Cat: 50926-M08H) from the overexpression lysate was verified.
A DNA sequence encoding the mouse SERPINI1 (O35684) (Met 1-Leu 410) was expressed, with a C-terminal polyhistidine tag.
The secreted recombinant mouse SERPINI1 comprises 405 amino acids and has a calculated molecular mass of 46 kDa. The apparent molecular mass of rmSERPINI1 is approximately 45-50 kDa in SDS-PAGE under reducing conditions as a result of glycosylation.
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.
Neuroserpin, also known as Protease inhibitor 12 and SERPINI1, is a secreted protein that belongs to the serpin family. Neuroserpin is a serine protease inhibitor that inhibits plasminogen activators and plasmin but not thrombin. Serine protease inhibitors of the serpin superfamily are involved in many cellular processes. Neuroserpin was first identified as a protein secreted from the axons of dorsal root ganglion neurons. Neuroserpin is predominantly expressed in the brain, and is expressed in the late stages of neurogenesis during the process of synapse formation. Overexpression of neuroserpin in an anterior pituitary corticotroph cell line results in the extension of neurite-like processes, suggesting that neuroserpin may play a role in cell communication, cell adhesion, and/or cell migration. Neuroserpin may be involved in the formation or reorganization of synaptic connections, as well as synaptic plasticity in the adult nervous system. Neuroserpin may also protect neurons from cell damage by tissue-type plasminogen activator. Defects of neuroserpin are the cause of familial encephalopathy with neuroserpin inclusion bodies (FEN1B).
serpin peptidase inhibitor, clade I (neuroserpin), member 1
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