Mouse IL5RA Baculovirus-Insect cells Overexpression Lysate

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Mouse IL5RA Baculovirus-Insect cells Overexpression Lysate: 产品信息

产品描述
This Mouse IL5RA overexpression lysate was created in Baculovirus-Insect cells and intented for use as a Western blot (WB) positive control. Purification of IL5RA protein (Cat: 51154-M08B) from the overexpression lysate was verified.
表达宿主
Baculovirus-Insect cells
种属
Mouse
蛋白构建信息
A DNA sequence encoding the mouse IL5Ra (P21183) (Met1-Val328) was expressed with a C-terminal polyhistidine tag.
分子量
The recombinant mouse IL5Ra comprises 321 amino acids and has a predicted molecular mass of 36.8 kDa. The apparent molecular mass of the protein is approximately 45 kDa in SDS-PAGE under reducing conditions due to glycosylation.

Mouse IL5RA Baculovirus-Insect cells 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 IL5RA Baculovirus-Insect cells Overexpression Lysate: 别称

Mouse CD125 Overexpression Lysate; Mouse CDw125 Overexpression Lysate; Mouse Il5r Overexpression Lysate

IL5RA 背景信息

Interleukin 5 receptor, alpha (IL5RA) also known as CD125 (Cluster of Differentiation 125) is a subunit of the Interleukin-5 receptor. IL5RA (CD125) is an interleukin 5 specific subunit of a heterodimeric cytokine receptor. The receptor is comprised of a ligand-specific alpha subunit and a signal transducing beta subunit shared by the receptors for interleukin 3 (IL3), colony-stimulating factor 2 (CSF2/GM-CSF), and interleukin 5 (IL5). The binding of this protein to IL5 depends on the beta subunit. The beta subunit is activated by the ligand binding and is required for the biological activities of IL5. This protein has been found to interact with syndecan binding protein (syntenin), which is required for IL5 mediated activation of the transcription factor SOX4. Six alternatively spliced transcript variants encoding three distinct isoforms have been reported. IL5RA (CD125) is a T-cell-derived cytokine that is particularly important in the development of asthma for the terminal differentiation, activation, and survival of committed eosinophil precursors.
全称
interleukin 5 receptor, alpha
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参考文献
  • Isobe M, et al. (1992) Localization of the gene encoding the alpha subunit of human interleukin-5 receptor (IL5RA) to chromosome region 3p24-3p26. Genomics. 14(3): 755-8.
  • Cheong HS, et al. (2005) Association analysis of interleukin 5 receptor alpha subunit (IL5RA) polymorphisms and asthma. J Hum Genet. 50(12): 628-34.
  • Isobe M, et al. (1992) Localization of the gene encoding the alpha subunit of human interleukin-5 receptor (IL5RA) to chromosome region 3p24-3p26. Genomics. 14(3): 755-8.
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