描述
CYTO-ID®自噬检测试剂盒 2.0
产品信息
品牌 |
产品名称 | 产品编号 | 规模 | 价格（元） |
| Ezno Life | CYTO-ID® Autophagy detection kit 2.0 CYTO-ID®自噬检测试剂盒 2.0 |
ENZ-KIT175-0200 | 200 tests | 7224.75 |
| Ezno Life | CYTO-ID® Autophagy detection kit 2.0 CYTO-ID®自噬检测试剂盒 2.0 |
ENZ-KIT175-0050 | 50 tests | 3420 |
产品特点：
- 无需转染检测，无需进行转染效率验证
- 更亮、更耐光的染料专门染色自噬囊泡
- 溶酶体染色可忽略不计，减少了其他染料所见的背景
- 快速量化天然异质细胞群中的自噬
- 促进自噬激活剂和抑制剂的高通量筛选
产品详情
| 应用： | 流式细胞术、荧光显微镜、荧光检测、HTS | |
| 应用说明： | CYTO-ID®自噬检测试剂盒 2.0 提供了一种快速、特异性和定量的方法，用于通过荧光显微镜、流式细胞术和酶标仪监测活细胞中的自噬。 | |
| 质量控制： | 显微镜检查：当 CYTO-ID® Green 自噬检测试剂 2 掺入细胞时，通常会在细胞核周区域的球形液泡中、分布在整个细胞质中的病灶或两个位置中观察到这种荧光探针的积累，具体取决于根据所研究的细胞类型。与用户手册中所述制备的自噬样品相比，未诱导的细胞显示出非常低的背景染色。 | |
| 数量： | 对于 -0200 尺寸： 200 次流式细胞术检测、250 次显微镜检测或 3 x 96 孔微孔板检测。对于 -0050 尺寸： 50 次流式细胞术检测、60 次显微镜检测或 1 x 96 孔微孔板检测。 |
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| 使用/稳定性： | 在妥善储存的情况下，试剂盒组件自收到之日起可稳定保存一年 | |
| 处理： | 避光。避免冷冻/解冻循环。 | |
| 船运： | 在蓝冰上发货 | |
| 短期储存： |
20°C |
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| 长期储存： | -80°C | |
| 内容： | CYTO-ID®绿色检测试剂 2 Hoechst 33342 核染色 自噬诱导剂（雷帕霉素） 氯喹对照 10X 检测缓冲液 |
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| 科学背景： |
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| 技术信息/产品说明： |
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| 协议： | 可在 bioprotocol.org 上找到原代 BMDC 中 FC 的详细协议：M. Stankov 等人在原代细胞中使用 CYTO-ID 染色对自噬活性进行流式细胞分析。 | |
| 监管状态： | RUO-仅供研究使用 |
产品参数：
CYTO-ID®自噬检测试剂盒 2.0 使用选择性标记积累的自噬泡的新型染料测量自噬泡并监测溶酶体抑制的活细胞中的自噬通量。该染料通过鉴定可滴定的功能部分进行了优化，这些部分允许溶酶体的染色最少，同时在掺入前自噬体、自噬体和自噬溶酶体（自噬溶酶体）时表现出明亮的荧光。该测定提供了一种无需细胞转染即可监测活细胞自噬的快速定量方法。
作用机制：
该探针是一种阳离子两亲示踪剂 (CAT) 染料，它以与诱导磷脂质沉着症的药物类似的方式迅速分配到细胞中。仔细选择染料上的可滴定功能部分可防止其在溶酶体中积累，但可以标记与自噬途径相关的液泡。
自噬示意图。细胞质物质被不断膨胀的膜囊（吞噬细胞）隔离，从而形成双膜囊泡，即自噬体。自噬体的外膜随后与溶酶体融合，内部物质在自噬溶酶体中降解。图中还描述了各种自噬调节剂。
基于流式细胞术的 Jurkat 细胞自噬分析。Jurkat 细胞未经处理或用 0.5μM 雷帕霉素 (RAP)、10μM 氯喹 (CLQ) 或两者处理 20 小时。用 CYTO-ID® Green Detection Reagent 2 染色 30 分钟后，洗涤细胞并通过流式细胞术进行分析。结果显示为直方图叠加。用 RAP + CLQ 处理的细胞显示荧光增加。
HeLa 细胞在 (A) 全培养基或 (B) 饥饿培养基 (EBSS) 和 40µM 氯喹中培养 4 小时后，用 CYTO-ID® Green 检测试剂 2 染色。在氯喹存在下在 EBSS 中饥饿的细胞显示出非常亮的绿色荧光信号和点状结构。
在 PBS 中测定 CYTO-ID® Green 检测试剂 2（图 A）的吸光度和荧光发射光谱 (499/548nm)。Hoechst 33342（图 B）的吸光度和荧光发射光谱 (350/461nm) 在 1X 检测缓冲液中测定。
基于微孔板的 HepG2 细胞自噬分析。在 DMSO（对照）、0.5 µM 雷帕霉素 (Rap)、10 µM 氯喹 (CLQ) 或 0.5 µM Rap 和 10 µM CLQ 中培养 20 小时后，HepG2 细胞被 CYTO-ID® Green Detection Reagent 2 染色。细胞也用 Hoechst 33342 染色以进行细胞数标准化。通过 CYTO-ID® Green Detection Reagent 2 测量，用雷帕霉素和氯喹处理的细胞自噬增加。
产品文献参考：
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