產(chǎn)品描述：

保存溫度：2-8度

主要優(yōu)點(diǎn)：

• 無需放射性物質(zhì)。
• 應(yīng)用 - 流式細(xì)胞儀或熒光顯微鏡。
• 檢測細(xì)胞水平的細(xì)胞溶解活性。
• 適用于多種類型的哺乳動(dòng)物細(xì)胞系。

技術(shù)

測量CMC / ADCCzui常用的方法是放射性鉻-51（51Cr）釋放試驗(yàn)（2）。該測定有幾個(gè)缺點(diǎn)：昂貴的，難以加載某些細(xì)胞類型，由于嚴(yán)格的環(huán)境規(guī)定而處置昂貴，并且具有51Cr自發(fā)釋放的高背景。使用流式細(xì)胞儀，現(xiàn)在可以消除對放射性物質(zhì)的需要，并增加在單個(gè)細(xì)胞基底上定量細(xì)胞溶解活性的能力。各組已經(jīng)證明通過流式細(xì)胞術(shù)測量CMC / ADCC活性與傳統(tǒng)的51Cr釋放測定（3,4,5,6）具有強(qiáng)烈（95％）的相關(guān)性。

測定原理

細(xì)胞跟蹤染料CFSE（7,8,9）用于標(biāo)記靶細(xì)胞群體。在測定完成實(shí)驗(yàn)方案后，加入7AAD（活/死）（10,11）以測量細(xì)胞死亡。7AAD僅進(jìn)入膜受損細(xì)胞并與DNA結(jié)合。

流式細(xì)胞術(shù)用于門靶細(xì)胞并測量7AAD陰性對7AAD后細(xì)胞。％細(xì)胞毒性通過以下等式計(jì)算（參見下面的實(shí)驗(yàn)例）：

7AAD正（右上象限）= R1 / 7AAD Postitve（右上象限）= R1 + 7AAD負(fù)（右下象限）= R2 ×100 
ACT1

為了測試豬gd淋巴細(xì)胞的自然殺傷能力，將K562細(xì)胞染色并調(diào)整至含有10％FBS的1×10 4個(gè)細(xì)胞/100μlPRMI的終濃度。以25:1,50:1和100:1的E / T比加入gd淋巴細(xì)胞，調(diào)節(jié)至400μlRPMI的總體積，然后在37℃，無菌封蓋的面管中孵育4小時(shí)。孵育后，將活/死染色劑直接加入每個(gè)管中，在冰上孵育15分鐘并通過流式細(xì)胞術(shù)分析。

引用文獻(xiàn)

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