Leuko Act Cktl With GolgiPlug 2x100uL

• 产品编号：BD-550583      品牌：BD-Pharmingen       原厂货号：550583
• 产品分类：生化试剂 > 免疫分析试剂 > 流式细胞术溶液和试剂
• 应用分类：

描述：

用于细胞内染色的细胞因子检测最关键的步骤是用于诱导产生细胞因子的细胞分离和活化。如果没有良好的活化细胞，培养时存在蛋白转运抑制剂，那么检测细胞因子细胞内积累就会受到影响。多细胞系激活素已专门应用于诱导产细胞因子的细胞。
 
激活素包含有：伴刀豆球蛋白A、脂多糖、佛波脂加钙离子或伊屋诺霉素、植物凝血素、葡萄球菌肠毒素B和TCR/CD3复合物亚基的单克隆抗体（含或不含有协同刺激素受体的抗体，比如CD28）。白细胞激活混合物，含BDA GolgiPlug™是一种即开可用的多细胞系活化混合物，包含佛波脂、PMA（12-十四酸佛波酯-13-乙酸盐）、钙离子（伊屋诺霉素）和蛋白转运抑制剂BD GolgiPlug;（布雷菲德菌素A）。这种混合物可用于引起T细胞响应的源发性细胞因子。使用这种混合物激活细胞产生细胞因子局限于细胞的糙面内质网。这种局限由蛋白转运抑制剂导致，布雷菲德菌素A可阻断细胞内蛋白转运。这样可提高细胞因子蛋白积累水平，增强荧光免疫检测和流式细胞分析性能。100uL成份足够处理50 ml细胞培养物（10^6个细胞/ml）因此，一个可刺激 ≥5 X 10^7 cells。

原厂资料：

The most critical step for detection of intracellular accumulations of cytokines by intracellular staining is the isolation and activation of a cell
population to induce production of the cytokine of interest. Without properly activated cytokine-producing cells, cultured in the presence of a
protein transport inhibitor, the ability to detect intracellular accumulations of cytokines in most cases is severely compromised. Various in
vitro methods have been reported for stimulating cells to produce cytokines. Polyclonal activators have been particularly useful for inducing
cytokine-producing cells. These activators include the following: concanavalin A, lipopolysaccharide, phorbol esters plus calcium ionophore
or ionomycin, phytohaemaglutinin, staphylococcus enterotoxin B, and monoclonal antibodies directed against subunits of the TCR/CD3
complex (with or without antibodies directed against costimulatory receptors, such as CD28).
The Leukocyte Activation Cocktail, with BD GolgiPlug™ is a ready -to-use polyclonal cell activation mixture containing the phorbol ester,
PMA (Phorbol 12-Myristate 13-Acetate), a calcium ionophore (Ionomycin) and the protein transport inhibitor BD GolgiPlug™ (Brefeldin A).
This mixture may be utilized to elicit a primary cytokine response from T cells. Stimulation of cells using this cocktail results in cytokine
production that is localized in the rough endoplasmic reticulum of cytokine -producing cells. This localization of cytokines is caused by the
protein transport inhibitor component, Brefeldin A which blocks intracellular protein transport processes. The increased accumulation of
cytokine protein levels in cells thus enhances detectability of cytokine-producing cells by immunofluorescent staining and flow cytometric
analysis. The contents of each 100 μL vial is sufficient for treating up to 50 mL of cell cultures (at ~10^6 cells/mL); therefore one can
stimulate ≥ 5 × 10^7 cells.

注意事项：

1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
2. Store product at -80°C prior to use or for long term storage of stock solutions.