Category
Neural Stem Cell Differentiation Markers
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/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.
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-H02H) 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 the Fc region of human IgG1 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