This Human ILT4 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of ILT4 protein (Cat: 14132-H02H) from the overexpression lysate was verified.
A DNA sequence encoding the human LILRB2 (AAH36827.1) (Met1-Val461) was expressed with the Fc region of human IgG1 at the C-terminus.
The recombinant human LILRB2/Fc is a disulfide-linked homodimer. The reduced monomer comprises 681 amino acids and has a predicted molecular mass of 74.8 kDa. The apparent molecular mass of the protein is approximately 93 kDa in SDS-PAGE under reducing conditions.
Human ILT4 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 ILT4 HEK293 Overexpression Lysate: 别称
Human CD85D Overexpression Lysate; Human ILT-4 Overexpression Lysate; Human ILT4 Overexpression Lysate; Human LIR-2 Overexpression Lysate; Human LIR2 Overexpression Lysate; Human MIR-10 Overexpression Lysate; Human MIR10 Overexpression Lysate
ILT4, also known as LILRB2, is a member of the subfamily B class of LIR receptors which contain two or four extracellular immunoglobulin domains, a transmembrane domain, and two to four cytoplasmic immunoreceptor tyrosine-based inhibitory motifs (ITIMs). ILT4 gene is a member of the leukocyte immunoglobulin-like receptor (LIR) family. Multiple transcript variants encoding different isoforms have been found for the ILT4 gene. ILT4 is expressed on immune cells where it binds to MHC class I molecules on antigen-presenting cells and transduces a negative signal that inhibits stimulation of an immune response. It is thought to control inflammatory responses and cytotoxicity to help focus the immune response and limit autoreactivity.