Mouse ICAM-1 HEK293 Overexpression Lysate

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Mouse ICAM-1 HEK293 Overexpression Lysate: 製品の情報

製品の説明
This Mouse ICAM-1 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of ICAM-1 protein (Cat: 50440-M03H) 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 extracellular domain of mouse ICAM1 (NP_034623.1) (Met 1-Asn 485) was fused with the C-terminal polyhistidine-tagged Fc region of human IgG1 at the C-terminus.
分子量
The recombinant mouse ICAM1/Fc is a disulfide-linked homodimer after removal of the signal peptide. The reduced monomer consists of 706 amino acids and has a predicted molecular mass of 78.3 kDa. In SDS-PAGE under reducing conditions, the apparent molecular mass of rm ICAM1/Fc monomer is approximately 120-130 kDa due to glycosylation.

Mouse ICAM-1 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 ICAM-1 HEK293 Overexpression Lysate: 別名

Mouse CD54 Overexpression Lysate; Mouse Icam-1 Overexpression Lysate; Mouse Ly-47 Overexpression Lysate; Mouse MALA-2 Overexpression Lysate

ICAM-1 背景情報

Intercellular adhesion molecule-1 (ICAM-1, or CD54) is a 9 kDa member of the immunoglobulin (Ig) superfamily and is critical for the firm arrest and transmigration of leukocytes out of blood vessels and into tissues. ICAM-1 is constitutively present on endothelial cells, but its expression is increased by proinflammatory cytokines. The endothelial expression of ICAM-1 is increased in atherosclerotic and transplant-associated atherosclerotic tissue and in animal models of atherosclerosis. Additionally, ICAM-1 has been implicated in the progression of autoimmune diseases. ICAM-1 is a ligand for LFA-1(integrin). When activated, leukocytes bind to endothelial cells via ICAM-1/LFA-1 interaction and then transmigrate into tissues. Presence with heavy glycosylation and other structural characteristics, ICAM-1 possesses binding sites for a number of immune-associated ligands and serves as the binding site for entry of the major group of human Rhinovirus (HRV) into various cell types. ICAM-1 also becomes known for its affinity for Plasmodium falciparum-infected erythrocytes (PFIE), providing more of a role in infectious disease. Previous studies have shown that ICAM-1 is involved in inflammatory reactions and that a defect in ICAM-1 gene inhibits allergic contact hypersensitivity.
完全な名称
intercellular adhesion molecule 1
研究分野
参考文献
  • Xu H, et al. (2001) The role of ICAM-1 molecule in the migration of Langerhans cells in the skin and regional lymph node. Eur J Immunol. 31(10): 3085-93.
  • Terol MJ, et al. (2003) Soluble intercellular adhesion molecule-1 (s-ICAM-1/s-CD54) in diffuse large B-cell lymphoma: association with clinical characteristics and outcome. Ann Oncol. 14(3): 467-74.
  • Mendez MP, et al. (2006) Shedding of soluble ICAM-1 into the alveolar space in murine models of acute lung injury. Am J Physiol Lung Cell Mol Physiol. 290(5): L962-70.
  • Lawson C, et al. (2009) ICAM-1 signaling in endothelial cells. Pharmacol Rep. 61(1): 22-32.
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