This Human CD58 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of CD58 protein (Cat: 12409-H02H) from the overexpression lysate was verified.
A DNA sequence encoding the human CD58 (Q9BRW0) (Met1-Arg215) was expressed with the Fc region of human IgG1 at the C-terminus.
The recombinant human CD58/Fc is a disulfide-linked homodimer. The reduced monomer comprises 428 amino acids and has a predicted molecular mass of 48.5 kDa. The apparent molecular mass of the protein is approximately 68 kDa in SDS-PAGE under reducing conditions.
Human CD58 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 CD58 HEK293 Overexpression Lysate: 别称
Human ag3 Overexpression Lysate; Human CD58 Overexpression Lysate; Human LFA-3 Overexpression Lysate; Human LFA3 Overexpression Lysate
CD53 is a member of the transmembrane 4 superfamily, also called the tetraspanin family. Most of these members are cell-surface proteins that are characterized by the presence of four hydrophobic domains. These proteins mediate signal transduction events that play a role in the regulation of cell development, activation, growth and motility. CD53 is a cell surface glycoprotein that is known to complex with integrins. Familial deficiency of CD53 gene has been linked to an immunodeficiency associated with recurrent infectious diseases caused by bacteria, fungi and viruses. CD53 contributes to the transduction of CD2-generated signals in T cells and natural killer cells and has been suggested to play a role in growth regulation.