Live or Dead 細胞活性檢測試劑盒是美國AAT Bioquest生產的細胞活性檢測試劑盒，AAT Bioquest的Mycolight 細菌活力測定試劑盒可在革蘭氏陽性和陰性細菌中提供細菌活力的雙色熒光測定。該試劑盒利用我們的綠色熒光核酸染色劑MycoLight Green和紅色熒光核酸染色碘化丙啶的混合物。單獨使用時，MycoLight 綠色污漬通常會標記群體中的所有細菌（活的和死的）。相比之下，碘化丙錠只穿透膜受損的細菌，當兩種染料都存在時，導致MycoLight 綠色染料熒光的減少。因此，使用MycoLight 綠色和碘化丙啶的適當混合物，具有完整細胞膜的活細菌發出綠色熒光，而具有損傷膜的死亡或瀕死的細菌 產生紅色熒光。Mycolight 細菌活力測定試劑盒是監測細菌種群活力的強大工具，可作為細胞膜完整性的函數。可以在510-530nm（FITC濾光片）和600-660nm（德克薩斯紅色濾光片光片）處以488nm處（常見的激發光源）激發熒光測定染色的細胞。百螢生物是AAT Bioquest的中國代理商，為您提供優質的Live or Dead 細胞活性檢測試劑盒。 

樣品實驗方案

簡要概述

1.用測試化合物制備細胞
2.添加染料加工溶液
3.在室溫或37°C孵育30分鐘至1小時
4.在Ex / Em = 610/650 nm（截止= 630 nm，紅色）和Ex / Em = 360/450 nm（截止= 420 nm，藍色）或在熒光顯微鏡下觀察（Texas red/Cy5通道   活細胞），（DAPI通道  死細胞）

工作溶液配制

將5μL的200X Cellbrite Red（組分A）和5μL的200X Nuclear Blue DCS1（組分C）加入1mL的測定緩沖液（組分B）中并充分混合以制備染料 - 工作溶液。 該染料加工溶液在室溫下穩定至少1小時。 注意：由于染色條件可能因細胞類型的不同而異，因此建議單獨確定組分A和C的適當濃度。有關細胞樣品制備的指南，請點擊查看。

操作步驟

1.根據標準方案準備細胞。 注意：我們用星形孢菌素（SS）在37℃下處理HeLa細胞4小時以誘導細胞凋亡。 詳細信息請參見圖1。

2.用100μL/孔（96孔板）或25μL/孔（384孔板）的染料加工溶液替換生長培養基。

3.將染料加工溶液板在室溫或37°C孵育30分鐘至1小時，避光。

4.用HHBS，PBS或您選擇的緩沖液洗滌細胞兩次。

5.向細胞中加入100μL/孔（96孔板）或25μL/孔（384孔板）的測定緩沖液（組分B）。

6.用熒光顯微鏡監測熒光信號，用德克薩斯紅或Cy5過濾器檢測活細胞，用DAPI過濾死細胞。 還可以使用熒光酶標儀（底部讀取模式）在Ex / Em = 610 / 650nm（截止= 630nm，紅色）和Ex / Em = 360 / 450nm（截止= 420nm，藍色）下分析熒光強度。

數據分析

參考文獻

Functional imaging of neuronal activity of auditory cortex by using Cal-520 in anesthetized and awake mice
Authors: Jingcheng Li, Jianxiong Zhang, Meng Wang, Junxia Pan, Xiaowei Chen, Xiang Liao
Journal: Biomedical Optics Express (2017): 2599--2610

NINJ2--A novel regulator of endothelial inflammation and activation
Authors: Jingjing Wang, Jingjing Fa, Pengyun Wang, Xinzhen Jia, Huixin Peng, Jing Chen, Yifan Wang, Chenhui Wang, Qiuyun Chen, Xin Tu
Journal: Cellular Signalling (2017)

Erythropoietin Stimulates Endothelial Progenitor Cells to Induce Endothelialization in an Aneurysm Neck After Coil Embolization by Modulating Vascular Endothelial Growth Factor
Authors: Peixi Liu, Yingjie Zhou, Qingzhu An, Yaying Song, Xi Chen, Guo-Yuan Yang, Wei Zhu
Journal: MEDICINE (2016): 1--8

Flexible Endoscopic Spray Application of Respiratory Epithelial Cells as Platform Technology to Apply Cells in Tubular Organs
Authors: Anja Lena Thiebes, Manuel Armin Reddemann, Johannes Palmer, Reinhold Kneer, Stefan Jockenhoevel, Christian Gabriel Cornelissen
Journal: Tissue Engineering Part C: Methods (2016): 322--331

Influence of hypothermia and subsequent rewarming upon leukocyte-endothelial interactions and expression of Junctional-Adhesion-Molecules A and B
Authors: Nicolai V Bogert, Isabella Werner, Angela Kornberger, Patrick Meybohm, Anton Moritz, Till Keller, Ulrich A Stock, Andres Beiras-Fernandez
Journal: Scientific reports (2016)

Inhibition of ABC transport proteins by oil sands process affected water
Authors: Hattan A Alharbi, David MV Saunders, Ahmed Al-Mousa, Jane Alcorn, Alberto S Pereira, Jonathan W Martin, John P Giesy, Steve B Wiseman
Journal: Aquatic Toxicology (2016): 81--88

Rapid generation of collagen-based microtissues to study cell--matrix interactions
Authors: Marie-Elena Brett, Alexandra L Crampton, David K Wood
Journal: Technology (2016): 1--8

Toxicokinetics and toxicodynamics of chlorpyrifos is altered in embryos of Japanese medaka exposed to oil sands process-affected water: evidence for inhibition of P-glycoprotein
Authors: Hattan A Alharbi, Jane Alcorn, Ahmed Al-Mousa, John P Giesy, Steve B Wiseman
Journal: Journal of Applied Toxicology (2016)

Spraying respiratory epithelial cells to coat tissue-engineered constructs
Authors: Anja Lena Thiebes, Stefanie Albers, Christian Klopsch, Stefan Jockenhoevel, Christian G Cornelissen
Journal: BioResearch open access (2015): 278--287

The efficient hemostatic effect of Antarctic krill chitosan is related to its hydration property
Authors: Shuai Wu, Zhuoyao Huang, Jianhui Yue, Di Liu, Ting Wang, Pierre Ezanno, Changshun Ruan, Xiaoli Zhao, William W Lu, Haobo Pan
Journal: Carbohydrate polymers (2015): 295--303

相關產品