Rhesus IL10RA HEK293 Overexpression Lysate

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Rhesus IL10RA HEK293 Overexpression Lysate: 产品信息

产品描述
This Rhesus IL10RA overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of IL10RA protein (Cat: 90125-C02H) from the overexpression lysate was verified.
表达宿主
HEK293 Cells
种属
Rhesus
蛋白构建信息
A DNA sequence encoding the rhesus IL10RA (XP_001092736.1) (Thr9-Asn245) was expressed, fused with the Fc region of human IgG1 at the C-terminus.
分子量
The recombinant rhesus IL10RA comprises 458 amino acids and has a calculated molecular mass of 51.8 KDa.

Rhesus IL10RA 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.

IL10RA 背景信息

IL10RA (Interleukin 10 Receptor Subunit Alpha, also known as CD210) is a Protein Coding gene. The protein encoded by this gene is a receptor for interleukin 10. Two transcript variants, one protein-coding, and the other, not protein-coding have been found for this gene. CD210 belongs to the type II cytokine receptor family. This protein is structurally related to interferon receptors. It has been shown to mediate the immunosuppressive signal of interleukin 10, and thus inhibits the synthesis of proinflammatory cytokines. Activation of CD210 leads to tyrosine phosphorylation of JAK1 and TYK2 kinases. CD210 is broadly expressed in the spleen, bone marrow, and other tissues. Diseases associated with IL10RA include Inflammatory Bowel Disease 28, Autosomal Recessive, and Inflammatory Bowel Disease 28.
全称
interleukin 10 receptor, alpha
研究领域
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    Jak-Stat Signaling Pathway
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    Autophagy Pathway
参考文献
  • Josephson K, et al. (2001) Purification, crystallization and preliminary X-ray diffraction of a complex between IL-10 and soluble IL-10R1. Acta Crystallogr D Biol Crystallogr. 57(Pt 12): 1908-11.
  • Tan, J C, et al. (1995) Characterization of recombinant extracellular domain of human interleukin-10 receptor. J Biol Chem. 270(21):12906-11.
  • Josephson, K, et al. (2001) Crystal structure of the IL-10/IL-10R1 complex reveals a shared receptor binding site. Immunity. 15(1):35-46.
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