Mouse CD73 HEK293 Overexpression Lysate

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Mouse CD73 HEK293 Overexpression Lysate: 产品信息

产品描述
This Mouse CD73 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of CD73 protein (Cat: 50231-M08H) from the overexpression lysate was verified.
表达宿主
HEK293 Cells
种属
Mouse
蛋白构建信息
A DNA sequence encoding the mouse NT5E (NP_035981.1) without the propeptide, corresponding to the amino acid (Met 1-Lys 549) was expressed, with a C-terminal polyhistidine tag.
分子量
The secreted recombinant mouse NT5E consists of 532 amino acids and has a calculated molecular mass of 59.4 kDa which is also detected in SDS-PAGE under reducing conditions.

Mouse CD73 HEK293 Overexpression Lysate: 使用指南

制备方法
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.
裂解缓冲液
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.
使用建议
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.
缓冲液
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.
应用
Western Blot (WB)
Optimal dilutions/concentrations should be determined by the end user.

Mouse CD73 HEK293 Overexpression Lysate: 别称

Mouse 2210401F01Rik Overexpression Lysate; Mouse AI447961 Overexpression Lysate; Mouse CD73 Overexpression Lysate; Mouse eNT Overexpression Lysate; Mouse NT Overexpression Lysate; Mouse Nt5 Overexpression Lysate

CD73 背景信息

5'-nucleotidase, also known as NT5E, NTE, and CD73, is a cell membrane protein that belongs to the 5'-nucleotidase family. CD73 is a glycosylphosphatidylinositol (GPI) anchored purine salvage enzyme expressed on the surface of human T and B lymphocytes. CD73 catalyzes the conversion of purine and pyrimidine ribo- and deoxyribonucleoside monophosphates to the corresponding nucleosides. CD73 serves as a costimulatory molecule in activating T cells. CD73 generated adenosine functions in cell signaling in many physiologic systems, including intestinal epithelium, ischemic myocardium, and cholinergic synapses. CD73 might mediate lymphocyte-stromal cell interactions or condition the local microenvironment to facilitate lymphocyte development and/or function. In CD73-depleted cells, surface levels of the leukocyte adhesion molecules ICAM-1, VCAM-1, and E-selectin increase. CD73 produces extracellular adenosine, which then acts on G protein-coupled purinergic receptors to induce cellular responses. CD73 has also been reported to regulate the expression of pro-inflammatory molecules in mouse endothelium.
全称
5'-nucleotidase, ecto (CD73)
参考文献
  • Resta R. et al., 1997, Cell Signal. 9 (2): 131-9.
  • Yamashita Y. et al., 1998, Eur J Immunol. 28 (10): 2981-90.
  • Louis NA. et al., 2008, J Immunol. 180 (6): 4246-55.
  • Grünewald JK. et al., 2010, J Inflamm. 7 (1): 10.
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