Mouse IL-12 HEK293 Overexpression Lysate

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Mouse IL-12 HEK293 Overexpression Lysate: 产品信息

产品描述
This Mouse IL-12 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of IL-12 protein (Cat: CT021-M02H) 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 p35 subunit of mouse IL12, termed as IL12A (P43431) (Met 1-Ala 215) was fused with the Fc region of human IgG1 at the C-terminus, constructed the plasmid 1; A DNA sequence encoding the p40 subunit of mouse IL12, termed as IL12B (P43432) (Met 1-Ser 335) was fused with the Fc region of human IgG1 at the C-terminus, constructed the plasmid 2. The two plasmids were co-expressed and the heterodimer was purified.
分子量
The recombinant mouse IL12 heterodimer of IL12A/IL12B, both was Fc chimera, comprises 988 (434 + 554) amino acids and has a calculated molecular mass of 111.5 (48.7 + 62.8) kDa. The apparent molecular mass of rh IL12 heterodimer is approximately 55 & 75 kDa respectively in SDS-PAGE under reducing conditions.

Mouse IL-12 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 IL-12 HEK293 Overexpression Lysate: 别称

Mouse Il-12a Overexpression Lysate; Mouse Il-12b Overexpression Lysate; Mouse IL-12p35 Overexpression Lysate; Mouse Il-12p40 Overexpression Lysate; Mouse Il12p40 Overexpression Lysate; Mouse Ll12a Overexpression Lysate; Mouse p35 Overexpression Lysate; Mouse p40 Overexpression Lysate

IL-12 背景信息

全称
interleukin 12A
Related Pathways
  • IL12 signaling pathway
    IL12 signaling pathway
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