1.百螢 鈣黃綠素紅鈉鹽參數(shù)：

E x (nm)? ? ?? ?562

E m (nm)? ? ? ?576

分??子 量? ? ? ?825.44

溶? ? ??劑? ? ? ??Water

存儲條件? ? ? ?在零下15度以下保存，避免光照

2.?百螢 鈣黃綠素紅鈉鹽簡介

? ? ? ?鈣黃綠素AM是最流行的熒光探針之一，用于標(biāo)記和檢測活細(xì)胞的細(xì)胞功能。但是，鈣黃綠素AM的單色使得無法在多色應(yīng)用中使用這種有價值的試劑。例如，由于與GFP相同的顏色，不可能在結(jié)合GFP的細(xì)胞中使用鈣黃綠素AM。為了解決鈣黃綠素AM的顏色限制，我們開發(fā)了鈣黃綠橙 ，鈣黃綠 和鈣黃綠 。這些新的Calcein AM類似物與Calcein AM結(jié)合使用，可以對活細(xì)胞進(jìn)行多色標(biāo)記和功能分析。非熒光鈣黃綠素 AM可以輕松進(jìn)入活細(xì)胞并水解產(chǎn)生強熒光鈣黃綠素 染料。可以使用通用的TRITC / Cy3濾波片組檢測Calcein Red 染料。

3.百螢 鈣黃綠素紅鈉鹽實驗方案

? ? ? ??AM酯是非極性酯，很容易穿過活細(xì)胞膜，并被活細(xì)胞內(nèi)的細(xì)胞酯酶迅速水解。AM酯被廣泛用于無創(chuàng)地將各種極性熒光探針裝載到活細(xì)胞中。但是，使用AM酯時必須特別注意，因為它們易于水解，特別是在溶液中。在使用高質(zhì)量無水二甲基亞砜（DMSO）之前，應(yīng)將其重新配制。DMSO儲備溶液可在-20°C的環(huán)境中干燥保存，并避光。在這些條件下，AM酯應(yīng)穩(wěn)定數(shù)月。

? ? ? ?以下是我們推薦的將AM酯加載到活細(xì)胞中的方案。該協(xié)議僅提供指南，應(yīng)根據(jù)您的特定需求進(jìn)行修改。

1）.準(zhǔn)備2至5 mM的AM酯高品質(zhì)無水DMSO儲備溶液。非離子型洗滌劑F-127有時用于增加AM酯的水溶性。

注：20％的Pluronic等體積??F-127溶液可以稀釋到裝載緩沖器之前加入到細(xì)胞活力指標(biāo)DMSO儲備溶液。終的Pluronic???F-127的濃度為約0.02％。

2）.在實驗當(dāng)天，將細(xì)胞活力的固體指示劑溶解在DMSO中，或?qū)⒅甘緞﹥淙芤旱牡确衷嚇咏鈨鲋潦覝?。在您選擇的緩沖液（例如Hanks和Hepes緩沖液）中準(zhǔn)備1至10 μM的工作溶液。對于大多數(shù)細(xì)胞系，建議終濃度為4至5 μM的細(xì)胞活力指標(biāo)。必須根據(jù)經(jīng)驗確定細(xì)胞加載所需指示劑的確切濃度。

3）.如果您的細(xì)胞中含有有機陰離子轉(zhuǎn)運蛋白，則可將丙磺舒（1-2.5 mM）或亞砜并吡嗪（0.1-0.25 mM）添加到細(xì)胞培養(yǎng)基中，以減少去酯化指示劑的泄漏。在室溫或37°C下將細(xì)胞與AM酯孵育20分鐘至一小時。

注意：降低裝載溫度可能會減少指示器的分隔。

4）.在無細(xì)胞活力指示劑的緩沖液（如果適用，包含陰離子轉(zhuǎn)運蛋白抑制劑）中洗滌細(xì)胞，以去除多余的探針。

注意：立即通過流式細(xì)胞儀分析細(xì)胞樣品，以測定零時每個細(xì)胞的平均熒光強度，無需清洗。

5）.在所需的Ex / Em波長下運行實驗。

4.參考文獻(xiàn)

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