Product Description
This Human DcR1 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of DcR1 protein (Cat: 10415-H08H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Sequence Information
A DNA sequence encoding the human TNFRSF10C (NP_003832.2) (Met1-Pro235) was expressed with a polyhistidine tag at the C-terminus.
Molecule Mass
The recombinant human TNFRSF10C consists of 221 amino acids and predicts a molecular mass of 23.6 kDa.
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).
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.
Application
Western Blot (WB)
Optimal dilutions/concentrations should be determined by the end user.
Human CD263 Overexpression Lysate;Human DCR1 Overexpression Lysate;Human DCR1-TNFR Overexpression Lysate;Human LIT Overexpression Lysate;Human TRAIL-R3 Overexpression Lysate;Human TRAILR3 Overexpression Lysate;Human TRID Overexpression Lysate
TNFRSF1C CNV in patients with CRC is associated with distant metastatic disease. A high frequency of CGI methylation in the TNFRSF1C promoter results in inactivation of the gene and enhancement of tumor growth in most PC cell lines (except CFPAC-1). Inactivation of TNFRSF1C by CpG island (CGI) hypermethylation can play an important role in PC progression and be potentially useful as a diagnostic marker and a new therapeutic approach for PC.