Mouse Semaphorin 5A HEK293 Overexpression Lysate: Product Information
This Mouse Semaphorin 5A overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of Semaphorin 5A protein (Cat: 50972-M08H) from the overexpression lysate was verified.
A DNA sequence encoding the mouse SEMA5A (Q3UPZ0) (Met1-Thr765) was expressed with a C-terminal polyhistidine tag.
The recombinant mouse SEMA5A comprises 755 amino acids and has a predicted molecular mass of 85.1 kDa. The apparent molecular mass of the protein is approximately 96 kDa in SDS-PAGE under reducing conditions due to 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.
Semaphorins are secreted, transmembrane, and GPI-linked proteins, defined by cysteine-rich semaphorin protein domains, that have important roles in a variety of tissues. Humans have 2 semaphorins, Drosophila has five, and two are known from DNA viruses. Semaphorins are found in nematodes and crustaceans but not in non-animals. They are grouped into eight classes on the basis of phylogenetic tree analyses and the presence of additional protein motifs. Semaphorins have been implicated in diverse developmental processes such as axon guidance during nervous system development and regulation of cell migration. Semaphorin-5A, also known as Semaphorin-F, Sema F, SEMA5A and SEMAF, is a single-pass type I membrane protein which belongs to thesemaphorin family. Semaphorin5A / SEMA5A contains onePSI domain, oneSema domain and sevenTSP type-1 domains. It may act as positive axonal guidance cues. Semaphorin5A / SEMA5A is an axon regulator molecule and plays major roles during neuronal and vascular development. It plays an essential role in embryonic development. Semaphorin5A / SEMA5A induces endothelial cell migration from pre-existing vessels. It also plays a role in autism, reducing the ability of neurons to form connections with other neurons in certain brain regions.
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