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蛋白转运抑制剂混合物 (500×)
有货
为什么选择此级别
BioReagent,无菌过滤 级 ，适用于对基线干扰要求严格的色谱和分析工作流程。
储存与运输
避光,-20°C储存。超低温运输 。请查阅批次 COA 获取详细规格。
质量文档
SDS、COA、产品数据表及规格说明书均可下载。可通过批号查询获取批次 COA。
文献证明
在色谱分析、有机合成和交叉偶联反应领域已被 0 篇同行评审文献引用。
概述
产品介绍
本产品为即用型蛋白转运抑制剂混合物（500X浓缩液），包含经浓度优化的布雷非德菌素A和莫能霉素钠。两者协同作用，可高效、特异地阻断细胞内高尔基体介导的蛋白质转运过程，广泛用于细胞内细胞因子、趋化因子等分泌蛋白的检测实验。本品与细胞刺激剂（如PMA/离子霉素）联用，可使新合成的目标蛋白在胞内有效累积，是进行流式细胞术胞内染色（ICS） 的关键试剂。
主要组分：
- 无水乙醇
- brefeldin A 1.5mg/mL，莫能霉素钠 0.7mg/mL
主要应用
- 流式细胞术胞内因子染色（ICS）：与细胞刺激混合物联用，用于检测T细胞、NK细胞等产生的细胞因子（如IFN-γ, TNF-α, IL-2, IL-4等）。
- 阻断蛋白质分泌：用于研究蛋白质的合成、加工与分泌过程。
- 适用于：悬浮细胞（如淋巴细胞）及贴壁细胞的胞内染色实验。
使用方法
标准使用流程
- 细胞刺激：在含有细胞的培养体系中，先加入刺激物（如抗原、PMA/离子霉素混合物）激活细胞。
- 添加抑制剂：在刺激开始的同时或稍后（通常推荐在刺激起始时加入），将本品按 1:500 的比例加入培养体系。例如，每1 mL细胞悬液加入 2 µL 本500X浓缩液。
- 孵育：将细胞置于37°C，5% CO₂培养箱中继续孵育。关键孵育时间：通常为4至6小时。此时间足以让细胞因子充分合成并在胞内累积，同时避免抑制剂毒性对细胞活力造成显著影响。最长不建议超过12小时。
- 后续处理：孵育结束后，收集细胞，按标准流程进行细胞表面染色、固定、破膜及胞内靶标染色。
注意事项
- 联用要求：本品为抑制剂，需与刺激物同时使用方有意义。单独使用仅会阻断基础水平的分泌。
- 时间控制：孵育时间是实验成功的关键。过短（<2小时）累积不充分，过长（>6小时）可能增加细胞非特异性染色背景并影响活力。
- 样品上清：使用本品后，细胞培养上清中无法检测到被阻断的分泌蛋白。如需分析分泌至培养基中的因子，请勿添加本品。
- 建议将产品分装保存，避免反复冻融。
Product Introduction
This product is a ready-to-use Protein Transport Inhibitor Cocktail (500X concentrate), containing optimized concentrations of Brefeldin A and Monensin Sodium. Working synergistically, they efficiently and specifically block the Golgi-mediated protein transport pathway within cells. It is widely used in intracellular detection assays for secretory proteins like cytokines and chemokines. When used in combination with cell stimulators (e.g., PMA/Ionomycin), it causes newly synthesized target proteins to accumulate intracellularly, making it a key reagent for Intracellular Cytokine Staining (ICS) by flow cytometry.
Key Components:
- Ethonal
- brefeldin A 1.5mg/mL，Monensin sodium salt 0.7mg/mL
Applications
- Co-stimulation Required: This is an inhibitor cocktail and must be used together with cell stimulators to be meaningful. Using it alone only blocks baseline secretion.
- Timing is Critical: Incubation time is key to success. Too short (<2 hours) leads to insufficient accumulation; too long (>6 hours) may increase non-specific background staining and affect viability.
- Supernatant Analysis: The secreted proteins blocked by this cocktail CANNOT be detected in the cell culture supernatant. Do not add this cocktail if you need to analyze factors secreted into the medium.
- Suitable for: Intracellular staining of both suspension cells (e.g., lymphocytes) and adherent cells.
Instructions for Use
Standard Protocol
- Cell Stimulation: Add stimulants (e.g., antigens, PMA/Ionomycin cocktail) to the cell culture to activate the cells.
- Add Inhibitors: At the beginning of stimulation (or shortly after), add this cocktail to the culture at a 1:500 dilution. For example, add 2 µL of the 500X concentrate to 1 mL of cell suspension.
- Incubation: Continue incubating the cells in a 37°C, 5% CO₂ incubator. Critical Incubation Time: Typically 4 to 6 hours. This allows sufficient protein synthesis and intracellular accumulation while minimizing the impact of inhibitor toxicity on cell viability. Incubation beyond 12 hours is not recommended.
- Post-Processing: After incubation, harvest the cells and proceed with standard protocols for surface staining, fixation, permeabilization, and intracellular staining.
Key Precautions
- Co-stimulation Required: This is an inhibitor cocktail and must be used together with cell stimulators to be meaningful. Using it alone only blocks baseline secretion.
- Timing is Critical: Incubation time is key to success. Too short (<2 hours) leads to insufficient accumulation; too long (>6 hours) may increase non-specific background staining and affect viability.
- Supernatant Analysis: The secreted proteins blocked by this cocktail CANNOT be detected in the cell culture supernatant. Do not add this cocktail if you need to analyze factors secreted into the medium.
Aliquot upon receipt to minimize freeze-thaw cycles.
规格
别名
流式专用蛋白转运抑制剂 | 流式专用蛋白转运抑制剂(布雷非德菌素A/莫能霉素混合物 500×) | 蛋白转运抑制剂混合物
英文别名
Flow Cytometry Protein Transport Inhibitor(Brefeldin A/Monensin Mixture) | Protein Transport Inhibitor Cocktail
规格或纯度
无菌过滤,BioReagent,500×
稳定性与储存
Aliquot upon receipt and store frozen at -20°C(12 months). Protect from light. Avoid repeated freeze-thaw cycles.
英文名称
Protein Transport Inhibitor Cocktail (500×)
储存条件
避光,-20°C储存
运输条件
超低温运输
产品线
生物产品
名称和识别符
分子类型
生物试剂/缓冲液
技术文档
高级数据
质检证书（CoA,COO,BSE/TSE 和分析图谱)
C of A & Other Certificates(BSE/TSE, COO):
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| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
 ZJ26F0434042 |
分析证书 | P1508367 |
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