Human IL-12 HEK293 Overexpression Lysate

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Human IL-12 HEK293 Overexpression Lysate: 产品信息

产品描述
This Human 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: GMP-CT011-H08H) 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 p35 subunit of human IL-12, termed as IL12A (P29459.2) (Met1-Ser219) was fused with a polyhistidine tag at the C-terminus, constructed the plasmid 1 ; A DNA sequence encoding the p40 subunit of human IL-12, termed as IL12B (NP_002178.2) (Met1-Ser328) was fused with a polyhistidine tag at the C-terminus, constructed the plasmid 2. The two plasmids were co-expressed and the heterodimer was purified.
分子量
The recombinant IL12 heterodimer of human IL12A/IL12B comprises 525 (208 + 317) amino acids and has a calculated molecular mass of 60.2 (24 + 36.2) kDa.

Human 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.

Human IL-12 HEK293 Overexpression Lysate: 别称

Human CLMF Overexpression Lysate; Human CLMF2 Overexpression Lysate; Human IL-12A Overexpression Lysate; Human IL-12B Overexpression Lysate; Human IL12 Overexpression Lysate; Human IL12 p70 Overexpression Lysate; Human IMD28 Overexpression Lysate; Human IMD29 Overexpression Lysate; Human NFSK Overexpression Lysate; Human NKSF Overexpression Lysate; Human NKSF1 Overexpression Lysate; Human NKSF2 Overexpression Lysate; Human P35 Overexpression Lysate

IL-12 背景信息

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