自噬(Autophagy )檢測 | Autophagy Cellular Analysis Kits 

CYTO-ID ^®自噬檢測試劑盒 |  Autophagy detection kit貨號ENZ-51031-K200

一種用於監測活細胞自噬的非轉染定量分析

自噬(Autophagy)是一種應激誘導的保護機制(stress-induced protective mechanism)。在基礎條件下不太活躍，真核細胞在受到某些不利條件（如營養耗盡和化學或環境壓力）時通過溶酶體介導(lysosome-mediated)的細胞內容物的大量降解來利用該機制。自噬活性(autophagic activity)增加在癌症(cancer)、神經變性(neurodegeneration)、心血管疾病( cardiovascular disease)和糖尿病( diabetes)的病理學中的作用已得到廣泛認可和普遍研究。如果溶酶體功能( lysosomal function)受到抑制，可以通過自噬小泡的增強積累來觀察自噬通量的誘導，從而阻止這些小泡的去除。

Enzo Life Sciences CYTO-ID ®自噬檢測試劑盒(CYTO-ID® Autophagy Detection Kit , ENZ-51031-K200)使用一種選擇性標記積累的自噬泡的新型染料測量自噬泡(autophagic vacuoles)並監測溶酶體抑制活細胞中的自噬流(autophagic flux )。488nm 可激發的綠色染料(488nm-excitable green dye )已通過鑑定可滴定功能部分進行了優化，這些部分允許對溶酶體的染色最少，同時在摻入前自噬體(pre-autophagosomes)、自噬體( autophagosomes)和自噬溶酶體（自噬溶酶體）時表現出明亮的螢光。該試劑盒還包括用於核染色的 Hoechst 33342 染料、自噬誘導劑-雷帕黴素(Rapamycin）和溶酶體抑制劑-氯喹（Chloroquine）。

作用機制 (Mechanism of Action & Autophagy Pathway)

CYTO-ID ^®自噬檢測試劑盒是 CELLESTIAL ^®產品線、試劑和檢測試劑盒的成員，其中包含螢光分子探針，這些探針已針對活細胞分析應用進行了廣泛的基準測試。該探針是一種陽離子兩親性示踪劑 (CAT) 染料，它以與誘導磷脂沉積症的藥物類似的方式快速分配到細胞中。仔細選擇染料上的可滴定功能部分可防止其在溶酶體中積累，但能夠標記與自噬途徑相關的液泡。

• 快速，無需轉染
• 使用已知的自噬活性抑制劑和激活劑驗證的方案
• 選擇性和全面的染色可以區分自噬通量和自噬溶酶體積累
• 溶酶體染色可忽略不計，降低了其他染料所見的背景
• 促進自噬激活劑和抑製劑的高通量篩選

快速，無需轉染 (Profile Autophagy Without Transfection)

Visualization of Autophagic Accumulation and Autophagic Flux 

無需轉染即可對自噬泡進行省時、快速和全面的標記。為了展示 CYTO-ID ^® Green 檢測試劑的優勢，首先用 RFP-LC3 表達載體轉染 HeLa 細胞，用 10 µM Tamoxifen 處理過夜，然後用 CYTO-ID ^® Green 檢測試劑染色。與基於過夜轉染的測定不同，CYTO-ID ^® Green 檢測試劑方法可在 15-30 分鐘內標記 100% 的細胞。圖 A：綠色信號表示自噬小泡的 CYTO-ID ^®綠色染色；圖 B：RFP-LC3 在成功轉染的細胞子集中的表達（紅色）；面板 C：合成圖像，顯示 CYTO-ID ^®綠色染料標記的囊泡與自噬體的特異性標記物 LC3 共定位。

Product Literature References
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