This Human TrkB overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of TrkB protein (Cat: 10047-H03H) from the overexpression lysate was verified.
A DNA sequence encoding the extracellular domain (Met 1-His 430) of human TrkB (NP_001007098.1) precursor was fused with the C-terminal polyhistidine-tagged Fc region of human IgG1 at the carboxy-terminus.
The recombinant human TrkB/Fc is a disulfide-linked homodimer. The reduced monomer consists of 646 amino acids and has a predicted molecular mass of 72 kDa. In SDS-PAGE under reducing conditions, the apparent molecular mass of rhTrkB/Fc monomer is approximately 110-120 kDa due to glycosylation.
Human TrkB 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.
1. Centrifuge the tube for a few seconds and ensure the pellet at the bottom of the tube.
2. Re-dissolve the pellet using 200μL pure water and boil for 2-5 min.
1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
稳定性 & 储存条件
Store at 4℃ for up to twelve months from date of receipt. After re-dissolution, aliquot and store at -80℃ for up to twelve months. Avoid repeated freeze-thaw cycles.
Western Blot (WB) Optimal dilutions/concentrations should be determined by the end user.
Human TrkB HEK293 Overexpression Lysate: 别称
Human GP145-TrkB Overexpression Lysate; Human GP145-TrkB/GP95-TrkB Overexpression Lysate; Human Tkrb Overexpression Lysate; Human trk-B Overexpression Lysate; Human TRKB Overexpression Lysate
TrkB receptor also known as TrkB tyrosine kinase or BDNF/NT-3 growth factors receptor or neurotrophic tyrosine kinase, receptor, type 2 (NTRK2) is a single transmembrane catalytic receptor with intracellular tyrosine kinase activity. TrkB/NTRK2 is a member of the neurotrophic tyrosine receptor kinase (NTRK) family. TrkB tyrosine kinase (TrkB) or NTRK2 is coupled to the Ras, Cdc42/Rac/RhoG, MAPK, PI3-K, and PLCgamma signaling pathways. There are four members of the Trk family; TrkA, TrkB, and TrkC and a related p75NTR receptor. Each family member binds different neurotrophins with varying affinities. TrkB/NTRK has the highest affinity for brain-derived neurotrophic factor (BDNF) and is involved in neuronal plasticity, long-term potentiation, and apoptosis of CNS neurons. Other neurotrophins include nerve growth factor(NGF), neurotrophin-3 and neurotrophin-4. TrkB/NTRK is a membrane-bound receptor that, upon neurotrophin binding, phosphorylates itself and members of the MAPK pathway. Signaling through this kinase leads to cell differentiation. Mutations in TrkB/NTRK have been associated with obesity and mood disorders.