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.
A DNA sequence encoding the human HCST (Q9UBK5-1) (Met-Pro48) was fused with Fc region of mouse IgG at the C-terminus.
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: 使用指南
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.
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.
1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
稳定性 & 储存条件
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.
Western Blot (WB) Optimal dilutions/concentrations should be determined by the end user.
Human DAP10 HEK293 Overexpression Lysate: 别称
Human DAP10 Overexpression Lysate; Human KAP10 Overexpression Lysate; Human PIK3AP Overexpression Lysate
HCST, also known as DAP1, belongs to the DAP1 family. It is a transmembrane signaling adaptor that contains a YxxM motif in its cytoplasmic domain. 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.