This Human TREM-2 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of TREM-2 protein (Cat: 11084-H08H) from the overexpression lysate was verified.
A DNA sequence encoding the extracellular domain of human TREM2 (NP_061838.1) (Met1-Ser174) was expressed fused with a polyhistidine tag at the C-terminus.
The secreted recombinant human TREM2 comprises 167 amino acids with a predicted molecular mass of 18.9 kDa. As a result of glycosylation, the apparent molecular mass of rhTREM2 is approximately 30-40 kDa band in SDS-PAGE under reducing conditions.
Human TREM-2 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 TREM-2 HEK293 Overexpression Lysate: 别称
Human TREM-2 Overexpression Lysate; Human Trem2a Overexpression Lysate; Human Trem2b Overexpression Lysate; Human Trem2c Overexpression Lysate
Triggering receptor expressed on myeloid cells 2 ( TREM2 ) is a single Ig domain receptor. It is expressed on macrophages and dendritic cells but not on granulocytes or monocytes. Its expression is most abundant in the basal ganglia, corpus callosum, medulla oblongata and spinal cord, and microglial cells are the major TREM2-producing cell type in the central nervous system (CNS). TREM2 may play a role in chronic inflammations and may stimulate production of constitutive rather than inflammatory chemokines and cytokines. TREM2 forms a receptor signaling complex with TYROBP and triggers activation of the immune responses in macrophages and dendritic cells. It also associates with the signal adapter protein, DAP12, which has a cytoplasmic ITAM, leading to the subsequent activation of cytoplasmic tyrosine kinases. TREM2 is both required and sufficient for competent uptake of apoptotic neuronal cells. TREM2 and TREM2-L form a receptor-ligand pair connecting microglia with apoptotic neurons, directing removal of damaged cells to allow repair. Deficiency of the adapter protein DAP12 or its associated receptor TREM2 is associated with abnormal osteoclast development in humans. Defects in TREM2 are causes of PLOSL, also known as NHD. In addition, TREM2 signaling is also an important pathway to promote healing of wounds in the colon where stem cell replacement is necessary.
triggering receptor expressed on myeloid cells 2
Bouchon, A. et al., 2000, J. Immunol. 164: 4991-4995.
Paloneva, J. et al., 2002, Am. J. Hum. Genet. 71:656-662.
Prada, I. et al., 2006, Neuroscience. 140 (4): 1139-48.
Neumann, H. et al., 2007, J Neuroimmunol. 184 (1-2): 92-9.
Thrash, JC. et al., 2009, Neurochem Res. 34 (1): 38-45.