Category
                                                                Neural Stem Cell Growth Factors
                             
                                                                        
                                                                                                                                                                
                                                                Product Name
                                                                Human NGFR/p75NTR HEK293 Overexpression Lysate
                             
                                                                        
                                                                        
                                                                        
                                                                Synonyms
                                                                Human CD271 Overexpression Lysate; Human Gp80-LNGFR Overexpression Lysate; Human p75(NTR) Overexpression Lysate; Human p75NTR Overexpression Lysate; Human TNFRSF16 Overexpression Lysate
                             
                                                                        
                                                                        
                                                                Molecule Mass
                                                                The secreted recombinant human NGFR consists of 233 amino acids and has a predicted molecular mass of 25 kDa. The apparent molecular mass of rhNGFR is approximately 45-60 kDa in SDS-PAGE under reducing conditions due to glycosylation.
                             
                                                                        
                                                                Stability & Storage
                                                                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.
                             
                                                                        
                                                                Product Description
                                                                This Human NGFR/p75NTR overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of NGFR/p75NTR protein (Cat: 13184-H08H) from the overexpression lysate was verified.
                             
                                                                        
                                                                Expression Host
                                                                HEK293 Cells
                             
                                                                        
                                                                Sequence Information
                                                                A DNA sequence encoding the human NGFR (P08138) extracellular domain (Met 1-Asn 250) was fused with a polyhistidine tag at the C-terminus.
                             
                                                                        
                                                                Preparation Method
                                                                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.
                             
                                                                        
                                                                Lysis Buffer
                                                                Modified RIPA Lysis Buffer: 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF.
                             
                                                                        
                                                                Recommend Usage
                                                                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.
                             
                                                                        
                                                                Sample Buffer
                                                                1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading  buffer).
                             
                                                                        
                                                                Application
                                                                Western Blot (WB)Optimal dilutions/concentrations should be determined by the end user.
                             
                                                                        
                                                                Related Pathways
                                                                Death Receptor Signaling