Rhesus CD200 HEK293 Overexpression Lysate

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Rhesus CD200 HEK293 Overexpression Lysate: 製品の情報

製品の説明
This Rhesus CD200 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of CD200 protein (Cat: 90230-C08H) from the overexpression lysate was verified.
発現ホスト
HEK293 Cells
Rhesus
タンパク質構築情報
A DNA sequence encoding the rhesus CD200 (NP_001244473.1) (Met1-Gly232) was expressed with a polyhistidine tag at the C-terminus.
分子量
The recombinant Rhesus CD200 comprises 213 amino acids and has a calculated molecular mass of 24 KDa. The apparent molecular mass of it is approximately 39-43 KDa respectively in SDS-PAGE.

Rhesus CD200 HEK293 Overexpression Lysate: 用法

調製方法
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.
溶解バッファー
Modified RIPA Lysis Buffer: 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF.
おすすめの用法
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.
バッファー
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.
アプリケーション
Western Blot (WB)
Optimal dilutions/concentrations should be determined by the end user.

CD200 背景情報

CD2 (OX-2) is a cell surface glycoprotein that imparts immune privileges by suppressing alloimmune and autoimmune responses through its receptor, CD2R, expressed primarily on myeloid cells. Signals delivered through the CD2:CD2R axis have been shown to play an important role in the regulation of anti-tumor immunity, and overexpression of CD2 has been reported in a number of malignancies, including CLL, as well as on cancer stem cells. The role of CD2-CD2R 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 CD2-CD2R, microglia activation, and Parkinson's disease (PD). The ability of CD2 to suppress myeloid cell activation is critical for maintaining normal tissue homeostasis but may also enhance the survival of migratory neoplastic cells. CD2 and CD2R associate via their respective N-terminal Ig-like domains. CD2 has been characterized as an important immunoregulatory molecule, increased expression of which can lead to decreased transplant rejection, autoimmunity, and allergic disease. Elevated CD2 expression has been reported to be associated with poor prognosis in a number of human malignancies. In addition, CD2 also plays an important role in prevention of graft rejection, autoimmune diseases and spontaneous abortion.
完全な名称
Cd200 molecule
参考文献
  • Minas K, et al. (2006) Is the CD200/CD200 receptor interaction more than just a myeloid cell inhibitory signal? Crit Rev Immunol. 26(3): 213-30.
  • Wang XJ, et al. (2007) CD200-CD200R regulation of microglia activation in the pathogenesis of Parkinson's disease. J Neuroimmune Pharmacol. 2(3): 259-64.
  • Wong KK, et al. (2010) The role of CD200 in immunity to B cell lymphoma. J Leukoc Biol. 88(2): 361-72.
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