Caspase-9活性檢測試劑盒

Caspase-9也稱ICE-LAP6或Mch6，可與細胞*素c和Apaf1形成復合物被激活，并進一步激活細胞凋亡的最關鍵酶caspase-3，從而觸發凋亡級聯反應。因此，Caspase-9是細胞凋亡信號轉導過程中重要的上游caspase。
細胞凋亡屬于程序化細胞死亡，可見于各種器官和細胞，在正常發育、生理、病理過程中對細胞和器官的穩態具有十分重要的調節作用。Caspase (Cysteine-requiring Aspartate Protease)是參與細胞凋亡過程的蛋白酶家族，包含10多個成員。Caspase-9也稱ICE-LAP6或Mch6，可與細胞*素c和Apaf1形成復合物被激活，并進一步激活細胞凋亡的最關鍵酶caspase-3，從而觸發凋亡級聯反應。因此，Caspase-9是細胞凋亡信號轉導過程中重要的上游caspase。其激活可以通過磷酸化進行調控。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。其吸光度值對應于Caspase-9的水解活性。反應體系100微升，試劑盒提供的試劑，除標準曲線外可進行50或100次樣品測定。適用：測定哺乳動物組織、細胞caspase-9活性。

組成與儲存：
1. 裂解緩沖液： 20 ml ，4 oC
2. 反應緩沖液： 10 ml ，4 oC
3. 2 mM Ac-LEHD-pNA底物： 0.5 ml，－20 oC避光
4. 10 mM pNA標準品： 0.5 ml，－20 oC避光
以上標出的量為100次檢測，50次檢測的試劑量減半。試劑常溫運輸。到達后按要求儲存，1年內穩定。

所需儀器：
酶標儀與96孔酶標板；或可見光分光光度計配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范圍內測定。

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Caspase-9活性檢測試劑盒 Caspase-9活性檢測試劑盒