Mouse B7-1 HEK293 Overexpression Lysate

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Mouse B7-1 HEK293 Overexpression Lysate: 製品の情報

製品の説明
This Mouse B7-1 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of B7-1 protein (Cat: 50446-M08H) 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 CD80 (Q00609-1) extracellular domain (Met 1-Lys 245) was expressed, with a polyhistidine tag at the C-terminus.
分子量
The recombinant mouse CD80 consists of 219 amino acids and has a predicted molecular mass of 25.1 kDa. In SDS-PAGE under reducing conditions, the apparent molecular mass of rm CD80 is approximately 48-50 kDa due to glycosylation.

Mouse B7-1 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 B7-1 HEK293 Overexpression Lysate: 別名

Mouse B71 Overexpression Lysate; Mouse Cd28l Overexpression Lysate; Mouse Ly-53 Overexpression Lysate; Mouse Ly53 Overexpression Lysate; Mouse MIC17 Overexpression Lysate; Mouse TSA1 Overexpression Lysate

B7-1 背景情報

The B-lymphocyte activation antigen B7-1 (referred to as B7), also known as CD8, is a member of cell surface immunoglobulin superfamily and is expressed on the surface of antigen-presenting cells including activated B cells, macrophages and dendritic cells. As costimulatory ligands, B7-1 which exists predominantly as dimer and the related protein B7-2, interact with the costimulatory receptors CD28 and cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) expressed on T cells, and thus constitute one of the dominant pathways that regulate T cell activation and tolerance, cytokine production, and the generation of CTL. The B7/CD28/CTLA4 pathway has the ability to both positively and negatively regulate immune responses. CD8 is thus regarded as promising therapeutic targets for autoimmune diseases and various carcinomas.
完全な名称
CD80 molecule
参考文献
  • Greenfield EA, et al. (1998) CD28/B7 costimulation: a review. Crit Rev Immunol. 18(5): 389-418.
  • Zang X, et al. (2007) The B7 family and cancer therapy: costimulation and coinhibition. Clin Cancer Res. 13(18 Pt 1): 5271-9.
  • Mir MA, et al. (2008) Signaling through CD80: an approach for treating lymphomas. Expert Opin Ther Targets. 12(8): 969-79.
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