Human DAP10 HEK293 Overexpression Lysate

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Human DAP10 HEK293 Overexpression Lysate: Product Information

Product Description
This Human DAP10 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of DAP10 protein (Cat: 14063-H05H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Species
Human
Sequence Information
A DNA sequence encoding the human HCST (Q9UBK5-1) (Met-Pro48) was fused with Fc region of mouse IgG at the C-terminus.
Molecule Mass
The recombinant human HCST/mFc is a disulfide-linked homodimer. The reduced monomer comprises 264 amino acids and has a predicted molecular mass of 29.3 kDa. The apparent molecular mass of the protein is approximately 34 in SDS-PAGE under reducing conditions.

Human DAP10 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.

Human DAP10 HEK293 Overexpression Lysate: Synonyms

Human DAP10 Overexpression Lysate; Human KAP10 Overexpression Lysate; Human PIK3AP Overexpression Lysate

DAP10 Background Information

HCST (Hematopoietic Cell Signal Transducer, also known as DAP1) is a Protein Coding gene. HCST belongs to the DAP10 family. It is a transmembrane signaling adaptor that contains a YxxM motif in its cytoplasmic domain. The encoded protein may form part of the immune recognition receptor complex with the C-type lectin-like receptor NKG2D. HCST may be part of the immune recognition receptor complex. This receptor plays a major role in triggering cytotoxicity against target cells expressing cell surface ligands such as MHC class I chain-related MICA and MICB, and UL16-binding proteins (ULBPs). It may activate phosphatidylinositol 3-kinase dependent signaling pathways through its intracytoplasmic YxxM motif.
Full Name
hematopoietic cell signal transducer
References
  • Wu J. et al., 1999, Science. 285 (5428): 730-2.
  • Karimi M. et al., 2006, J Immunol. 175 (12): 7819-28.
  • André P. et al., 2004, Eur J Immunol. 34 (4): 961-71.
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