This Human DC-SIGN overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of DC-SIGN protein (Cat: 10200-H01H) from the overexpression lysate was verified.
A DNA sequence encoding the N-terminally truncated extracellular domain (Lys 62-Ala 404) of human DC-SIGN (NP_066978.1) was expressed with the fused Fc region of human IgG1 at the N-terminus.
The recombinant human Fc/DC-SIGN chimera is a disulfide-linked homodimeric protein. The reduced monomer consists of 580 amino acids and has a calculated molecular mass of 65.8 kDa. As a result of glycosylation, the apparent molecular mass of rh Fc/DC-SIGN is approximately 75 kDa in SDS-PAGE under reducing conditions.
Human DC-SIGN 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 DC-SIGN HEK293 Overexpression Lysate: 別名
Human CD209 Overexpression Lysate; Human CDSIGN Overexpression Lysate; Human CLEC4L Overexpression Lysate; Human DC-SIGN Overexpression Lysate; Human DC-SIGN1 Overexpression Lysate; Human MGC129965 Overexpression Lysate
Dendritic cell (DC)-specific intercellular adhesion molecule 3 (ICAM-3) grabbing nonintegrin (DC-SIGN), also known as CD29, is a type II transmembrane protein on DCs with a C-type lectin extracellular domain, is capable of binding ICAM-3 on resting T cells in the secondary lymphoid organs, providing the initial contact between these cells during the establishment of cell-mediated immunity. It is not only a pattern recognition receptor but implicated in immunoregulation of DCs. It has important role in mediating DC adhesion, migration, inflammation, activating primary T cell, triggering immune response and participating in immune escape of pathogens and tumors. DC-SIGN also mediates capture and internalization of viral, bacterial, and fungal pathogens by dendritic cells, such as HIV-1, Ebola virus, cytomegalovirus, Dengue virus, and hepatitis C virus. DC-SIGN is unique in that it regulates adhesion processes, such as DC trafficking and T-cell synapse formation, as well as antigen capture. Moreover, even though several C-type lectins have been shown to bind HIV-1, DC-SIGN does not only capture HIV-1 but also protects it in early endosomes allowing HIV-1 transport by DC to lymphoid tissues, where it enhances trans infection of T cells.