This Human TIE2 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of TIE2 protein (Cat: 10700-H08H) from the overexpression lysate was verified.
A DNA sequence encoding the extracellular domain (Met 1-Lys 745) of human Tie2 (NP_000450.2) precursor was fused with a polyhistidine tag at the C-terminus.
The recombinant human Tie2 comprises 734 amino acids and predicts a molecular mass of 82 kDa. As a result of glycosylation, the apparent molecular mass of rhTie2 is approximately 95-105 kDa in SDS-PAGE under reducing conditions.
Human TIE2 HEK293 Overexpression Lysate: 用法
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).
安定性 & 保存条件
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 TIE2 HEK293 Overexpression Lysate: 別名
Human CD202B Overexpression Lysate; Human TIE-2 Overexpression Lysate; Human TIE2 Overexpression Lysate; Human VMCM Overexpression Lysate; Human VMCM1 Overexpression Lysate
TEK, or TIE-2, is an endothelial cell-specific receptor tyrosine kinase (RTK) that is known as a functioning molecule of vascular endothelial cells. TEK comprises a subfamily of RTK with TIE, and these two receptors play critical roles in vascular maturation, maintenance of integrity and remodeling. Targeted mutagenesis of both Tek and its agonistic ligand, Angiopoietin-1, result in embryonic lethality, demonstrating that the signal transduction pathways mediated by this receptor are crucial for normal embryonic development. TEK signaling is indispensable for the development of the embryonic vasculature and suggests that TEK signaling may also be required for the development of the tumor vasculature.