Mouse ICOS HEK293 Overexpression Lysate

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Mouse ICOS HEK293 Overexpression Lysate: 製品の情報

製品の説明
This Mouse ICOS overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of ICOS protein (Cat: 50466-M02H) from the overexpression lysate was verified.
発現ホスト
HEK293 Cells
Mouse
タンパク質構築情報
A DNA sequence encoding the mouse ICOS (Q9WVS0) extracellular domain (Met 1-Leu 142) was fused with the Fc region of human IgG1 at the C-terminus.
分子量
The recombinant mouse ICOS/Fc is a disulfide-linked homodimer. The reduced monomer consists of 363 amino acids and has a predicted molecular mass of 40.9 kDa. In SDS-PAGE under reducing conditions, the apparent molecular mass of rmICOS/Fc monomer is approximately 40-45 kDa.

Mouse ICOS HEK293 Overexpression Lysate: 用法

調製方法
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.
溶解バッファー
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.
おすすめの用法
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.
バッファー
1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
安定性 & 保存条件
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.
アプリケーション
Western Blot (WB)
Optimal dilutions/concentrations should be determined by the end user.

Mouse ICOS HEK293 Overexpression Lysate: 別名

Mouse AILIM Overexpression Lysate; Mouse CCLP Overexpression Lysate; Mouse CRP-1 Overexpression Lysate; Mouse H4 Overexpression Lysate; Mouse Ly115 Overexpression Lysate

ICOS 背景情報

Inducible costimulator (ICOS), also called AILIM (activiation-inducible lymphocyte immunomediatory molecule) is a cell-surface receptor, and belongs to the CD28 family of immune costimulatory receptors consisting of CD28, CTLA-4 and PD-1. The interaction of B7-H2/ICOS plays a critical role in Th cell differentiation, T−B cell interactions which is essential for germinal center formation, and humoral immune responses, and as well as the production of cytokine IL-4. In addition, ICOS is more potent in the induction of IL-1 production, a cytokine important for suppressive function of T regulatory cells. The B7-1/B7-2--CD28/CTLA-4 and ICOS-B7RP-1 pathway provides key second signals that can regulate the activation, inhibition and fine-tuning of T-lymphocyte responses. ICOS stimulates both Th1 and Th2 cytokine production but may have a preferential role in Th2 cell development. Moreover, The B7-1/B7-2-CD28/CTLA-4 and ICOS-B7RP-1 pathway has been suggested of being involved in the development of airway inflammation and airway hyperresponsiveness.
完全な名称
inducible T-cell co-stimulator
参考文献
  • Coyle AJ, et al. (2004) The role of ICOS and other costimulatory molecules in allergy and asthma. Springer Semin Immunopathol. 25(3-4): 349-59.
  • Chen YQ, et al. (2006) CD28/CTLA-4--CD80/CD86 and ICOS--B7RP-1 costimulatory pathway in bronchial asthma. Allergy. 61(1): 15-26.
  • van Berkel ME, et al. (2006) CD28 and ICOS: similar or separate costimulators of T cells Immunol Lett. 105(2): 115-22.
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