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偵測細胞膜的改變                                                                                                              偵測粒線體膜電位改變




    偵測細胞膜的改變
                                                            細胞凋亡不會引起發炎反應，因為在晚期細胞會形成                                                【Mitochondrial Membrane Potential Kit】SI-MAK159 / SI-MAK160
                                                            凋亡小體讓巨噬細胞、或是自噬細胞所吞食。研究發
                                                            現細胞凋亡時期，細胞膜會產生一些變化，這些細胞                                                  偵測原理
                                                            膜的變化可能是一些“訊息”；讓體內啟動自我排除機                                                  細胞凋亡初期，線粒體轉膜電位會發生變化，                        實驗操作流程
                                                            制。細胞凋亡時細胞膜變化有以下幾個特徵                                                       導致膜穿透性的改變。利用是陽離子染劑JC-
                                                            1. 膜表面糖蛋白的sialic acid residues消失                                          10，可以偵測到細胞膜的通透改變。JC-10在
                                                            2. 細胞失去原本細胞膜的不對稱性；例如：內膜的                                                  正常細胞中，會在線粒體中形成聚集體，發出
                                                               物質Phosphatidylserine暴露至外膜。                                             強烈的紅色螢光。但是在凋亡的細胞中，因線

                                                            因此可利用對Phosphatidylserine具有高度親和性的                                          粒體穿膜電位的改變，會以單體形式存在細胞
                                                            Annexin-V，去偵測細胞膜的變化。                                                      液中，而發出綠色螢光。



                      偵 測 方 法         相 關 產 品                        產 品 編 號                   包 裝
                        Flow          Annexin-V-FLUOS                ROC-11828681001          250Test                                                                             Add 50ul of JC-10 dye ,
                                                                                                                                                                                  and incubate 30~60 min.
                     螢光顯微鏡            Annexin-V-FLUOS Staining Kit   ROC-11858777001          50Test                                                                              Then add 50 ul of assay
                                                                                                                                                                                  buffer B
                                                                                                                                                                                                            Monitor the fluorescence
                     螢光顯微鏡            Annexin-V-FITC kit             XR-LAVK050               50 Test                                                                                                       intensity (lex = 490/
                        Flow                                         XR-LAVK250               250Test                                                                                                       lem = 525 nm) and (lex =
                                                                                                                                                                                                            540/lem = 590 nm) for
                                                                                                                                                                                                            ratio analysis. The ratio
                                                                                                                                                                                                            of red/green fluorescence
                                                                                                                                                                                                            intensity is used to
               【 Annexin-V-FLUOS / Annexin-V-FLUOS Staining Kit 】                                                                                                                                           determine MMP.
                                                    ROC-11828681001/ ROC-11858777001

                偵測原理與標的                        實驗的操作流程
                利用帶有螢光染料的AnnexinV、
                去結合凋亡細胞；再搭配PI與
                BOBI兩種螢光染劑區別細胞凋亡
                與細胞壞死。

                適用的實驗分析方法
                --Flow
                --螢光顯微鏡
                                                                                                                                          Campotothecin-induced mitochondria membrane potential   Effect of FCCP induced mitochondria membrane
                                                                                                                                          changes were measured with JC-10 and JC-1 in Jurkat   potential change in JurKat cells. JurKat cells were
                適用的偵測檢體                                                                                                                   cells. After Jurkat cells were treated with camptothecin   dye loaded with JC-10 dye-loading solution along
                --細胞(懸浮or貼附)                   區別正常細胞，凋亡細胞與壞死細胞的染色結果                                                                     (10 μM) for 4 hours, JC-1 and JC-10 dye loading   with DMSO (Top) or 5 μM FCCP (Low) for 10
                                                                                                                                                                                          minutes. The fluorescent intensities for both J-
                                                                                                                                          solutions were added to the wells and incubated for 30
                --組織切片                                                 Normal     Apoptotic     Necrotic                                  minutes. The fluorescent intensities for both J-aggregates   aggregates and monomeric forms of JC-10 were
                                                                       cells      cells         cells                                     and monomeric forms of JC-1 and JC-10 were measured   measured with a flow cytometer using FL1 and FL2
                                                                                                                                                                                          channels. Uncompensated data (left column) were
                                                                                                                                          at Ex/Em = 490/525 nm and 490/590 nm.
                產品優點                               Annexin-V staining      -           +            +                                                                                     compared with compensated data (right column).
                • 操作快速：只需15分鐘染色時間
                • 可快速分辨凋亡的細胞                       Propidium iodide        -           -            +                                    相 關 產 品                          偵 測 方 式                 產 品 品 號            包 裝
                                                   staining
                • 可使用Flow 或 顯微鏡觀察
                • 可以區別細胞凋亡或細胞壞死                    BOBO-1                  -           -            +                                    Mitochondria Membrane Potential   Microplate Readers    SI-MAK159          500Test
                                                                                                                                         Kit (JC-10 assay)                Flow                   SI-MAK160          100Test







     UNI-ONWARD                                             4
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