This Mouse TrkC overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of TrkC protein (Cat: 50320-M08H) from the overexpression lysate was verified.
A DNA sequence encoding the extracellular domain of mouse TrkC (NP_032772.3) (Met 1-Thr 429) was expressed, with a C-terminal polyhistidine tag.
The secreted recombinant mouse TrkC comprises 409 amino acids and has a calculated molecular mass of 46.1 kDa. As a result of glycosylation, the recombinant protein migrates as an approximately 100-110 kDa band in SDS-PAGE under reducing conditions.
Mouse TrkC 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.
NT-3 growth factor receptor is also known as neurotrophic tyrosine kinase receptor type 3 or TrkC tyrosine kinase or Trk-C receptor, is a member of the neurotrophic tyrosine receptor kinase (NTRK) family. This kinase is a membrane-bound receptor that, upon neurotrophin binding, phosphorylates itself and members of the MAPK pathway. TrkC/NTRK3 is widely expressed in the developing and adult nervous system. In later embryonic development, TrkC/NTRK3 is expressed in various structures of the CNS including the caudate-putamen, septal nuclei, cerebellum, and brainstem. Other neurotrophins include nerve growth factor(NGF), neurotrophin-3 and neurotrophin-4. In the PNS, The trkC hybridization appears to correlate, both temporally and spatially, with the outgrowth of axons toward their peripheral targets. TrkC/NTRK3 is widely expressed in the three identified branches of the mammalian nervous system and appears to correlate with the expression of NT-3, its cognate ligand. The apparent colocalization of trkC transcripts with NT-3 raises the possibility this neurotrophin exerts its trophic effects by a paracrine and/or autocrine mechanism. Signaling through this kinase leads to cell differentiation and may play a role in the development of proprioceptive neurons that sense body position. Mutations in the TrkC encoding gene have been associated with medulloblastomas, secretory breast carcinomas, and other cancers.