Human ICOS ligand HEK293 Overexpression Lysate

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Human ICOS ligand HEK293 Overexpression Lysate: 产品信息

产品描述
This Human ICOS ligand overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of ICOS ligand protein (Cat: 11559-H02H) 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 human ICOSLG (O75144-1) (Met1-Ser258) was expressed with the Fc region of human IgG1 at the C-terminus.
分子量
The recombinant human ICOSLG/Fc is a disulfide-linked homodimer. The reduced monomer comprises 481 amino acids and has a predicted molecular mass of 53.7 kDa. The apparent molecular mass of the protein is approximately 66-76 and 33 kDa in SDS-PAGE under reducing conditions.

Human ICOS ligand 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 ligand HEK293 Overexpression Lysate: 别称

Human B7-h2 Overexpression Lysate; Human B7H2 Overexpression Lysate; Human B7RP-1 Overexpression Lysate; Human B7RP1 Overexpression Lysate; Human CD275 Overexpression Lysate; Human GL50 Overexpression Lysate; Human ICOS ligand Overexpression Lysate; Human ICOS-L Overexpression Lysate; Human ICOSL Overexpression Lysate; Human ICOSLG Overexpression Lysate; Human LICOS Overexpression Lysate

ICOS ligand 背景信息

Inducible co-stimulator ligand (ICOSL), also known as B7-H2, is a member of the B7 family of co-stimulatory molecules related to B7-1 and B7-2. It is a transmembrane glycoprotein with extracellular IgV and IgC domains and binds to ICOS on activated T cells, thus delivers a positive costimulatory signal for optimal T cell function. The structural features of ICOSL are crucial for its costimulatory function. The present study shows that ICOSL displays a marked oligomerization potential, resembling more like B7-1 than B7-2. B7-H2-dependent signaling may play an active role in a proliferative response rather than in cytokine and chemokine production. The CD28/B7 and ICOS/B7-H2 pathways are both critical for costimulating T cell immune responses. Deficiency in either pathway results in defective T cell activation, cytokine production, and germinal center formation.
全称
inducible T-cell co-stimulator ligand
研究领域
参考文献
  • Flesch IE. (2002) Inducible costimulator-ligand (ICOS-L). J Biol Regul Homeost Agents. 16(3): 217-9.
  • Kajiwara K, et al. (2009) Expression and function of the inducible costimulator ligand B7-H2 in human airway smooth muscle cells. Allergol Int. 58(4): 573-83.
  • Wong SC, et al. (2009) Functional hierarchy and relative contribution of the CD28/B7 and ICOS/B7-H2 costimulatory pathways to T cell-mediated delayed-type hypersensitivity. Cell Immunol. 256(1-2): 64-71.
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