Human ICOS HEK293 Overexpression Lysate

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Human ICOS HEK293 Overexpression Lysate: 产品信息

产品描述
This Human ICOS overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of ICOS protein (Cat: 10344-H03H) 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 extracellular domain of human ICOS (NP_036224.1) (Met 1-Phe 141) was fused to the C-terminal polyhistidine-tagged Fc region of human IgG1 at the C-terminus.
分子量
The recombinant human ICOS/Fc is a disulfide-linked homodimeric protein. The reduced monomer consists of 368 amino acids and predicts a molecular mass of 41.6 kDa. The apparent molecular mass of rhICOS/Fc monomer is approximately 50 kDa due to glycosylation.

Human ICOS 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 ICOS HEK293 Overexpression Lysate: 别称

Human AILIM Overexpression Lysate; Human CD278 Overexpression Lysate; Human CVID1 Overexpression Lysate

ICOS 背景信息

Inducible costimulator (ICOS), also called AILIM (Activation-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 are essential for the germinal center formation, and humoral immune responses, and as well as the production of cytokine IL-4. Also, ICOS is more potent in the induction of IL-10 production, a cytokine important for the suppressive function of T regulatory cells. The B7-1/B7-2--CD28/CTLA-4 and ICOS-B7RP-1 pathway provide 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 as being involved in the development of airway inflammation and airway hyperresponsiveness.
全称
inducible T-cell co-stimulator
研究领域
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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