Mouse DR5/TRAIL R2 HEK293 Overexpression Lysate

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Mouse DR5/TRAIL R2 HEK293 Overexpression Lysate: Product Information

Product Description
This Mouse DR5/TRAIL R2 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of DR5/TRAIL R2 protein (Cat: 50412-M08H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Species
Mouse
Sequence Information
A DNA sequence encoding the extracellular domain of mouse TNFRSF10B (NP_064671.2) (Met 1-Ser 177) was expressed, with a C-terminal polyhistidine tag.
Molecule Mass
The secreted recombinant mouse TNFRSF10B consists of 136 amino acids and has a calculated molecular mass of 15 kDa. As a result of glycosylation, the apparent molecular mass of the recombinant protein is approximately 25-35 kDa in SDS-PAGE under reducing conditions.

Mouse DR5/TRAIL R2 HEK293 Overexpression Lysate: Usage Guide

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.

Mouse DR5/TRAIL R2 HEK293 Overexpression Lysate: Synonyms

Mouse DR5 Overexpression Lysate; Mouse KILLER Overexpression Lysate; Mouse Ly98 Overexpression Lysate; Mouse MK Overexpression Lysate; Mouse TRAILR2 Overexpression Lysate; Mouse TRICK2A Overexpression Lysate; Mouse TRICK2B Overexpression Lysate; Mouse TRICKB Overexpression Lysate

DR5/TRAIL R2 Background Information

Tumor necrosis factor receptor superfamily, member 1b, official symbol TNFRSF1B, also known as Death receptor 5, CD262, TNF-related apoptosis-inducing ligand receptor 2 (TRAIL R2), is a member of the TNF-receptor superfamily, and contains an intracellular death domain. This receptor can be activated by tumor necrosis factor-related apoptosis inducing ligand (TNFSF1/TRAIL/APO-2L), and transduces an apoptosis signal. Studies with FADD-deficient mice suggested that FADD, a death domain containing adaptor protein, is required for the apoptosis mediated by this protein. TRAIL R2/CD262/TNFRSF1B was purified independently as the only receptor for TRAIL detectable on the surface of two different human cell lines that undergo apoptosis upon stimulation with TRAIL. TRAIL R2/CD262/TNFRSF1B contains two extracellular cysteine-rich repeats, typical for TNF receptor (TNFR) family members, and a cytoplasmic death domain. TRAIL R2/CD262/TNFRSF1B mediates apoptosis via the intracellular adaptor molecule FADD/MORT1. TRAIL receptors can signal both death and gene transcription, functions reminiscent of those of TNFR1 and TRAMP, two other members of the death receptor family. Defects in TRAIL R2/CD262/TNFRSF1B may be a cause of head and neck squamous cell carcinomas (HNSCC) also known as squamous cell carcinoma of the head and neck.
Full Name
tumor necrosis factor receptor superfamily, member 10b
Research Areas
Related Pathways
  • Death Receptor Signaling
    Death Receptor Signaling
  • p53 Pathway
    p53 Pathway
References
  • Schneider P, et al. (1997) TRAIL receptors 1 (DR4) and 2 (DR5) signal FADD-dependent apoptosis and activate NF-kappaB. Immunity. 7(6): 831-6.
  • Ichikawa K, et al. (2003) TRAIL-R2 (DR5) mediates apoptosis of synovial fibroblasts in rheumatoid arthritis. J Immunol. 171(2): 1061-9.
  • Walczak H, et al. (1997) TRAIL-R2: a novel apoptosis-mediating receptor for TRAIL. EMBO J. 16(17): 5386-97.
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