This Human CD200 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of CD200 protein (Cat: 10886-H03H) from the overexpression lysate was verified.
A DNA sequence encoding the human CD200 (NP_005935.4) extracellular domain (Met 1-Gly 232) was fused with the C-terminal polyhistidine-tagged Fc region of human IgG1 at the C-terminus.
The recombinant human CD200/Fc is a disulfide-linked homodimer. The reduced monomer consists of 450 amino acids and has a predicted molecular mass of 50.5 kDa. As a result of glycosylation, the apparent molecular mass of rh CD200/Fc monomer is approximately 65-70 kDa in SDS-PAGE under reducing conditions.
Human CD200 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 CD200 HEK293 Overexpression Lysate: 别称
Human MOX1 Overexpression Lysate; Human MOX2 Overexpression Lysate; Human MRC Overexpression Lysate; Human OX-2 Overexpression Lysate; Human OX2 Overexpression Lysate
CD200 (OX-2) is a cell surface glycoprotein that imparts immune privileges by suppressing alloimmune and autoimmune responses through its receptor, CD200R, expressed primarily on myeloid cells. Signals delivered through the CD200:CD200R axis have been shown to play an important role in the regulation of anti-tumor immunity, and overexpression of CD200 has been reported in a number of malignancies, including CLL, as well as on cancer stem cells. The role of CD200-CD200R signaling in immune regulation of the central nervous system has become a popular field of research in recent years. Many studies have shown that there is a close correlation between CD200-CD200R, microglia activation, and Parkinson's disease (PD). The ability of CD200 to suppress myeloid cell activation is critical for maintaining normal tissue homeostasis but may also enhance the survival of migratory neoplastic cells. CD200 and CD200R associate via their respective N-terminal Ig-like domains. CD200 has been characterized as an important immunoregulatory molecule, increased expression of which can lead to decreased transplant rejection, autoimmunity, and allergic disease. Elevated CD200 expression has been reported to be associated with poor prognosis in some human malignancies. Besides, CD200 also plays an important role in prevention of graft rejection, autoimmune diseases and spontaneous abortion.