This Human NGFR overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of NGFR protein (Cat: 13184-H02H) from the overexpression lysate was verified.
A DNA sequence encoding the human NGFR (P08138) extracellular domain (Met 1-Asn 250) was fused with the Fc region of human IgG1 at the C-terminus.
The secreted recombinant human NGFR/Fc is a disulfide-linked homodimeric protein. The reduced monomer consists of 463 amino acids and has a predicted molecular mass of 50.6 kDa. The apparent molecular mass of rhNGFR/Fc monomer is approximately 70-80 kDa in SDS-PAGE under reducing conditions due to glycosylation.
Human NGFR 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 NGFR HEK293 Overexpression Lysate: 别称
Human CD271 Overexpression Lysate; Human Gp80-LNGFR Overexpression Lysate; Human p75(NTR) Overexpression Lysate; Human p75NTR Overexpression Lysate; Human TNFRSF16 Overexpression Lysate
Nerve growth factor receptors (NGFRs) belong to a large growth factor receptor family. NGFR includes two types of receptors: high-affinity nerve growth factor receptor and low-affinity nerve growth factor receptor. The high-affinity nerve growth factor receptor is also referred to as the Trk family whose members are bound by some neurotrophins with high affinity. Nerve growth factor binds with TrkA after being released from target cells, the NGF / TrkA complex is subsequently trafficked back to the cell body. The Low-affinity nerve growth factor receptor also named p75 which binds with all kinds of neurotrophins with low affinity. All four kinds of neurotrophins, including Nerve growth factor, Brain-derived neurotrophic factor, Neurotrophin-3, and Neurotrophin-4 bind to the p75. Studies have proved that NGFR acts as a molecular signal switch that determines cell death or survival by three steps. First, pro-nerve growth factor (prNGF) triggers cell apoptosis by its high-affinity binding to p75NTR, while NGF induces neuronal survival with low-affinity binding. Second, p75NTR mediates cell death by combining with co-receptor Sortilin, whereas it promotes neuronal survival through combination with proNGF. Third, the release of the intracellular domain chopper or cleavage short p75 NTR can independently initiate neuronal apoptosis.
nerve growth factor receptor (TNFR superfamily, member 16)
Death Receptor Signaling
Chen LW, et al. (2008) The proNGF-p75NTR-sortilin signalling complex as new target for the therapeutic treatment of Parkinson's disease. CNS Neurol Disord Drug Targets. 7(6): 512-23.
Deponti D, et al. (2009) The low-affinity receptor for neurotrophins p75NTR plays a key role for satellite cell function in muscle repair acting via RhoA. Mol Biol Cell.20(16): 3620-7.
Ken-ichiro K, et al. (2004) Necdin-related MAGE proteins differentially interact with the E2F1 transcription factor and the p75 neurotrophin receptor. J Biol Chem. 279 (3): 1703-12.