Human CD3E & CD3G Heterodimer HEK293 Overexpression Lysate

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Human CD3E & CD3G Heterodimer HEK293 Overexpression Lysate: 产品信息

产品描述
This Human CD3E & CD3G Heterodimer overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of CD3E & CD3G Heterodimer protein (Cat: CT041-H0305H) from the overexpression lysate was verified.
表达宿主
HEK293 Cells
种属
Human
蛋白构建信息
A DNA sequence encoding the extracellular domain (Met1-Asp126) of human CD3E (NP_000724.1) was fused with the C-terminal FLAG-tagged Fc region of human IgG1 at the C-terminus, constructed the plasmid 1; A DNA sequence encoding the extracellular domain (Met1-Ser116) of human CD3G (NP_000064.1) was fused the Fc region of mouse IgG1 at the C-terminus, constructed the plasmid 2. The two plasmids were co-expressed and the human CD3E/CD3G heterodimer was purified.
分子量
The recombinant heterodimer of human CD3E/CD3G comprises 681 (353+328) amino acids and has a calculated molecular mass of 76.9 (39.8+37.1) KDa. As a result of glycosylation, the apparent molecular mass of human CD3E/CD3G heterodimer is approximately 42-47 KDa in SDS-PAGE under reducing conditions.

Human CD3E & CD3G Heterodimer 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.

Human CD3E & CD3G Heterodimer HEK293 Overexpression Lysate: 别称

Human CD3E & CD3G Heterodimer Overexpression Lysate; Human IMD18 Overexpression Lysate; Human T3E Overexpression Lysate; Human TCRE Overexpression Lysate

CD3E & CD3G Heterodimer 背景信息

全称
CD3e molecule, epsilon (CD3-TCR complex)
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