Abbkine 活細胞和死細胞雙染色試劑盒 | 貨號KTA1001
Abbkine 活細胞和死細胞雙染色試劑盒 | 貨號KTA1001
細胞介導細胞毒性(Cell-mediated cytotoxicity)是免疫系統(immune system) 對體內受損細胞，cytolysis的一個重要現象。區分活細胞和死細胞對於研究生長控制和細胞死亡具重要性。
Fig.1. Hela cells stained with Abbkine Live and Dead Cell Double Staining Kit. A: Live cells stain green, B: Dead cells (30min incubation in 70% ethanol) stain red.
Fig.2. Hela cells incubated with staining solution in Abbkine Live and Dead Cell Double Staining Kit for 20min.
產品描述： Abbkine 活細胞和死細胞雙染色試劑盒 (Abbkine Live and Dead Cell Double Staining Kit) 提供了一個方便的測定,進行評估細胞的可行性；套組提供兩種probes同時測定活細胞和死細胞, 可用於測量細胞健康的識別參數:血漿膜完整性(plasma membrane integrity) 和細胞內酯酶活性(intracellular esterase activity)。該試劑盒利用LiveDye活細胞染劑; 一種細胞滲透的綠色螢光染料(cell-permeable green fluorescent dye , Ex/Em = 488/530 nm)染色活細胞(live cells)。NucleiDye核染料,為一種細胞不可滲透的紅色螢光染料(Ex/Em = 535/617) , 用於染色死細胞(dead cells)。
- Kit components
- LiveDye
• NucleiDye
• Assay Buffer (10X) - Features & Benefits
- Suitable for mammalian cells staining.
• Useful for the rapid quantitation of cell viability using flow cytometery or fluorescent microscopy.
• LiveDye (green, Ex/Em=488/530nm), NucleiDye (red, Ex/Em= 535/617nm). - Usage notes
NucleiDye is a potential mutagen. Use appropriate precautions when handling this reagent.
- Storage instructions
Refer to list of materials supplied for storage conditions of individual components. Stable for at least 12 months at recommended temperature from date of shipment.
- Shipping
Gel pack with blue ice.
引用文獻
- Ocular surface repair using decellularized porcine conjunctivaL Zhao, Y Jia, C Zhao. Acta biomaterialia, 2019 – Elsevier
- Grafting antibacterial polymer brushes from titanium surface via polydopamine chemistry and activators regenerated by electron transfer ATRPShuangshuang Wang, Jixuan Song, Yuchao Li, et al. Reactive and Functional Polymers. July 2019, 140: 48-55.
實驗程序 (標準流程見原廠網站)
注意：核染料的最佳濃度和反應時間，依不同實驗應用或細胞類型需再最佳化。
1.處理細胞依操作用手冊建議方法。
2.對於非貼壁細胞，通過離心（4oC，600g，5min）收集1-2×106個細胞。對於貼壁細胞，首先用胰蛋白酶(Trypsin)消化細胞，然後離心。
3.用冰冷的PBS洗滌細胞沉澱兩次。
4.將細胞600g離心5分鐘，然後將試管轉移至冰中。
5.用冰冷的70％乙醇(70% ethanol)在蒸餾水中緩慢重懸細胞。將細胞放在-20°C持續2小時或更長時間（建議隔夜固定12-24小時）。在染色和分析之前，細胞可以在-20°C下保存數週。
注意：將細胞固定在單個細胞懸液中非常重要。細胞固定時容易結塊。非常緩慢地逐滴添加初始體積的70％乙醇同時輕輕渦旋有助於防止細胞結塊。
6.在4°C下以1000g離心5分鐘。
7.除去乙醇，並將細胞重懸於1 mL冰冷的PBS中。
8.在4°C下以500g將細胞離心10分鐘，然後除去PBS。重複步驟7和8兩次徹底除去乙醇。
9.將細胞重懸於0.5 mL染色溶液中，並於37°C孵育30分鐘 (在黑暗)。
10.用PBS洗滌細胞兩次，並用冰冷的PBS重懸。分析其中的細胞（Ex / Em = 535/615 nm）通過流式細胞儀檢測 (24小時內)。
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