Caspase-9也稱ICE-LAP6或Mch6，可與細(xì)胞*素c和Apaf1形成復(fù)合物被激活，并進(jìn)一步激活細(xì)胞凋亡的最關(guān)鍵酶caspase-3，從而觸發(fā)凋亡級聯(lián)反應(yīng)。因此，Caspase-9是細(xì)胞凋亡信號轉(zhuǎn)導(dǎo)過程中重要的上游caspase。
細(xì)胞凋亡屬于程序化細(xì)胞死亡，可見于各種器官和細(xì)胞，在正常發(fā)育、生理、病理過程中對細(xì)胞和器官的穩(wěn)態(tài)具有十分重要的調(diào)節(jié)作用。Caspase (Cysteine-requiring Aspartate Protease)是參與細(xì)胞凋亡過程的蛋白酶家族，包含10多個(gè)成員。Caspase-9也稱ICE-LAP6或Mch6，可與細(xì)胞*素c和Apaf1形成復(fù)合物被激活，并進(jìn)一步激活細(xì)胞凋亡的最關(guān)鍵酶caspase-3，從而觸發(fā)凋亡級聯(lián)反應(yīng)。因此，Caspase-9是細(xì)胞凋亡信號轉(zhuǎn)導(dǎo)過程中重要的上游caspase。其激活可以通過磷酸化進(jìn)行調(diào)控。試劑盒測定原理基于Caspase-9特異水解其多肽底物Ac-LEHD-pNA (N-acetyl-Leu-Glu-His-Asp-p-nitroanilide)，釋放出游離的硝基ben胺pNA，后者呈黃色在405 nm具有最大吸收峰，用可見光分光光度比色方法測定。其吸光度值對應(yīng)于Caspase-9的水解活性。反應(yīng)體系100微升，試劑盒提供的試劑，除標(biāo)準(zhǔn)曲線外可進(jìn)行50或100次樣品測定。適用：測定哺乳動(dòng)物組織、細(xì)胞caspase-9活性。

組成與儲存：
1. 裂解緩沖液： 20 ml ，4 oC
2. 反應(yīng)緩沖液： 10 ml ，4 oC
3. 2 mM Ac-LEHD-pNA底物： 0.5 ml，－20 oC避光
4. 10 mM pNA標(biāo)準(zhǔn)品： 0.5 ml，－20 oC避光
以上標(biāo)出的量為100次檢測，50次檢測的試劑量減半。試劑常溫運(yùn)輸。到達(dá)后按要求儲存，1年內(nèi)穩(wěn)定。

所需儀器：
酶標(biāo)儀與96孔酶標(biāo)板；或可見光分光光度計(jì)配100μl容積的石英比色杯。
工作波長：最大吸收波長405 nm，如不具備也可在400~450 nm范圍內(nèi)測定。

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Caspase-9活性檢測試劑盒 Caspase-9活性檢測試劑盒