Annexin V (細胞表面 PS 分析)-Cell Biology-盟基生物科技股份有限公司

Apoptosis (細胞凋亡)

Annexin V (細胞表面 PS 分析)

Annexin V (細胞表面 PS 分析)

Apoptosis (細胞凋亡)

Annexin V (細胞表面 PS 分析)

細胞凋亡實驗中,偵測細胞膜表面 Phosphatidylserine (PS),超多種染劑選擇。

細胞凋亡 (apoptosis) 為程序性的細胞死亡 (programmed cell death),由身體內或外的刺激所導致。細胞凋亡為生物體重要的生理作用,和胚胎時期的發育以及成熟體中的組織更新、免疫系統、生殖系統皆有關係。整個過程由基因所調控,導致 DNA 裂解、細胞膜變形、形成凋亡小體 (apoptotic body) 等。研究中常透過四種方法檢測細胞凋亡的早期到晚期,分別為 Annexin V apoptosis assay (細胞表面 PS 分析)、Caspase assay (凋亡蛋白酶分析)、Mitochondrial membrane potential assay (粒線體膜電位分析)、TUNEL assay (斷裂 DNA 分析) (表一)。
細胞膜50%皆由膜脂質構成,膜的內外分布不同的脂質,其中 Phosphatidylserine (PS) (磷脂醯絲胺酸) 只存在於細胞膜內側。當細胞凋亡發生時,PS 會翻轉到細胞膜外面。Annexin V 為 Ca2+ 磷脂質結合蛋白,和 PS 具有高親合力。因此,透過螢光標記的 Annexin V,搭配流式細胞儀分析,可以專一性地偵測細胞膜外側有無 PS。Elabscience 提供傳統染劑標記 Annexin V,Biotium 則提供創新 CF® Dye 染劑標記 Annexin V。


表一:細胞凋亡的四種檢測方法比較
方法 Annexin V apoptosis assay
(細胞表面 PS 分析)
Caspase assay
(凋亡蛋白酶分析)
Mitochondrial membrane potential assay
(粒線體膜電位分析)
TUNEL assay
(斷裂 DNA 分析)
偵測目標 Phosphatidylserine (PS) Caspase activity Mitochondrial membrane potential Fragmented DNA
凋亡階段 Early --> --> -->Late
偵測方法 螢光 螢光、吸光值 螢光 螢光
反應時間 15-30 mins 螢光:15-30 mins
吸光值:2-4 hours
15 mins 60-120 mins
 
一、產品介紹
Elabscience 提供 Biotin、傳統螢光染劑、螢光蛋白標記的 Annexin V,和 Propidium Iodide (PI) 或 7-AAD 的細胞凋亡檢測套組。Propidium Iodide (PI) 和 7-AAD 為核酸 DNA 染劑,無法穿透細胞膜,因此只會染色死細胞。透過同時染色 Annexin V 和 Propidium Iodide (PI) 或 7-AAD,可同時檢測細胞凋亡和死亡 (細胞凋亡晚期) 的細胞族群 (圖一)。除了套組之外,Elabscience 也提供單獨 Annexin V 試劑,包含液體和 Azide-free 的凍乾形式。Azide-free 的凍乾形式不含有防腐劑,對細胞或動物傷害更小,保存也更穩定。
圖一:Jurkat 細胞經過5 μM Camptothecin 處理4小時 (左) 或未處理 (右),並以 Annexin V-FITC 和 Propidium Iodide (PI) 染色,觀察細胞凋亡。經過 Camptothecin 處理後,細胞凋亡的細胞族群大幅度地增加。

Biotium 則提供 CF® Dye 染劑標記的 Annexin V,有多種螢光選擇,包括 Near-IR 螢光。CF® Dye 為創新染劑,比起一般的染劑更強和更穩定。除了液體之外,Biotium 還提供 Azide-free 的凍乾形式,不含有防腐劑,對細胞或動物傷害更小,保存也更穩定。並且,Biotium 也有單獨販售 7-AAD 和 Propidium Iodide (PI) (三、產品資訊:表六)。
圖二:Jurkat 細胞未處理 (DMSO)(blue) 或以0.1uM, 0.25uM, 1uM staurosporine (red) 處理4小時,並以 NucView®530 和 CF405M® Annexin V 檢測。可以明顯地看出隨著藥物處理濃度增加,細胞凋亡的族群也顯著性地增加。


二、產品優點
1. 套組好方便:
Elabscience 提供多種 Annexin V 套組選擇,搭配 7-AAD 或 Propidium Iodide (PI) (三、產品資訊:表二)。

2. 超多種螢光選擇
Biotium 提供創新19種 CF® Dye 螢光選擇,Elabscience 則提供 Biotin 和14種傳統螢光染劑和螢光蛋白選擇。


三、產品資訊

表二:Annexin V 套組訂購資訊  
貨號 產品名稱 Annexin V 螢光 Ex/Em 包裝
30060 CF®488A and 7-AAD Apoptosis Assay Kit CF®488A: 490/515 nm 100 assays
30061 CF®488A and PI Apoptosis Assay Kit
E-CK-A211 Annexin V-FITC / PI Apoptosis Detection Kit FITC: 490/530 nm 20T/50T/100T/200T
E-CK-A212 Annexin V-FITC / 7-AAD Apoptosis Detection Kit
E-CK-A213 Annexin V-AF647 / PI Apoptosis Detection Kit AF647: 650/670 nm
E-CK-A214 Annexin V-AF647 / 7-AAD Apoptosis Detection Kit
E-CK-A216 Annexin V-PE / 7-AAD Apoptosis Detection Kit PE: 495;565/575 nm
E-CK-A217 Annexin V-APC / PI Apoptosis Detection Kit APC: 650/660 nm
E-CK-A218 Annexin V-APC / 7-AAD Apoptosis Detection Kit
E-CK-A219 Annexin V-EGFP / PI Apoptosis Detection Kit EGFP: 488/510 nm
E-CK-A220 Annexin V-EGFP / 7-AAD Apoptosis Detection Kit
E-CK-A221 Annexin V-Cyanine5 / PI Apoptosis Detection Kit Cyanine5: 650/670 nm
E-CK-A222 Annexin V-Cyanine5 / 7-AAD Apoptosis Detection Kit
E-CK-A228 Annexin V-PE/Cyanine7 / 7-AAD Apoptosis Detection Kit PE/Cyanine7: 495;565;755/775 nm
E-CK-A229 Annexin V-APC/Cyanine7 / PI Apoptosis Detection Kit APC/Cyanine7: 650;760/780 nm
E-CK-A230 Annexin V-APC/Cyanine7 / 7-AAD Apoptosis Detection Kit
E-CK-A233 Annexin V-EV450 / PI Apoptosis Detection Kit EV450: 410/450 nm
E-CK-A234 Annexin V-EV450 / 7-AAD Apoptosis Detection Kit
E-CK-A235 Annexin V-EV500 / PI Apoptosis Detection Kit EV500: 425/500 nm
E-CK-A236 Annexin V-EV500 / 7-AAD Apoptosis Detection Kit
E-CK-A237 Annexin V-AF488 / PI Apoptosis Detection Kit AF488: 495/520 nm
E-CK-A238 Annexin V-AF488 / 7-AAD Apoptosis Detection Kit
 
表三:CF®Dye 鍵結 Annexin V 單獨試劑訂購資訊  
 
貨號 產品名稱 螢光 Ex/Em 包裝
29012 CF®350 Annexin V 347/448 nm 0.5 mL (50 ug/mL)
29012R-5ug CF®350 Annexin V, Azide-Free, Lyophilized 5ug
29009 CF®405M Annexin V 408/452 nm 0.5 mL (50 ug/mL)
29009R-5ug CF®405M Annexin V, Azide-Free, Lyophilized 5ug
29083 CF®450 Annexin V 450/538 nm 0.5 mL (50 ug/mL)
29083R-5ug CF®450 Annexin V, Azide-Free, Lyophilized 5ug
29005 CF®488A Annexin V 490/515 nm 0.5 mL (50 ug/mL)
29005R-5ug CF®488A Annexin V, Azide-Free, Lyophilized 5ug
29004 CF®555 Annexin V 555/565 nm 0.5 mL (50 ug/mL)
29004R-5ug CF®555 Annexin V, Azide-Free, Lyophilized 5ug
29010 CF®568 Annexin V 562/583 nm 0.5 mL (50 ug/mL)
29010R-5ug CF®568 Annexin V, Azide-Free, Lyophilized 5ug
29011 CF®594 Annexin V 593/614 nm 0.5 mL (50 ug/mL)
29011R-5ug CF®594 Annexin V, Azide-Free, Lyophilized 5ug
29085 CF®583R Annexin V 586/609 nm 0.5 mL (50 ug/mL)
29085R-5ug CF®583R Annexin V, Azide-Free, Lyophilized 5ug
29008 CF®633 Annexin V 630/650 nm 0.5 mL (50 ug/mL)
29008R-5ug CF®633 Annexin V, Azide-Free, Lyophilized 5ug
29014 CF®640R Annexin V 642/662 nm 0.5 mL (50 ug/mL)
29014R-5ug CF®640R Annexin V, Azide-Free, Lyophilized 5ug
29003 CF®647 Annexin V 650/665 nm 0.5 mL (50 ug/mL)
29003R-5ug CF®647 Annexin V, Azide-Free, Lyophilized 5ug
29069 CF®660R Annexin V 663/682 nm 0.5 mL (50 ug/mL)
29069R-5ug CF®660R Annexin V, Azide-Free, Lyophilized 5ug
29007 CF®680 Annexin V, Azide-Free, Lyophilized 681/698 nm 25 ug
29070 CF®680R Annexin V, Azide-Free, Lyophilized 680/701 nm 25 ug
29082 CF®700 Annexin V, Azide-Free, Lyophilized 699/737 nm 25 ug
29006 CF®750 Annexin V, Azide-Free, Lyophilized 755/777 nm 25 ug
29046 CF®770 Annexin V, Azide-Free, Lyophilized 770/797 nm 25 ug
29047 CF®790 Annexin V, Azide-Free, Lyophilized 784/806 nm 25 ug
29078 CF®800 Annexin V, Azide-Free, Lyophilized 797/816 nm 25 ug
 
表四:其他螢光染劑鍵結 Annexin V 單獨試劑訂購資訊
貨號 產品名稱 螢光 Ex/Em 包裝
E-CK-A110 Annexin V-Biotin Reagent x 50T/100T/200T/500T
E-CK-A110U Annexin V-Biotin Azide-Free Lyophilized Powder 25ug/50ug/100ug/200ug
E-CK-A111 Annexin V-FITC Reagent 490/530 nm 50T/100T/200T/500T
E-CK-A111U Annexin V-FITC Azide-Free Lyophilized Powder 25ug/50ug/100ug/200ug
E-CK-A113 Annexin V-AF647 Reagent 650/670 nm 50T/100T/200T/500T
E-CK-A113U Annexin V-AF647 Azide-Free Lyophilized Powder 25ug/50ug/100ug/200ug
E-CK-A115 Annexin V-PE Reagent 495;565/575 nm 50T/100T/200T/500T
E-CK-A117 Annexin V-APC Reagent 650/660 nm 50T/100T/200T/500T
E-CK-A119 Annexin V-EGFP Reagent 488/510 nm 50T/100T/200T/500T
E-CK-A119U Annexin V-EGFP Azide-Free Lyophilized Powder 25ug/50ug/100ug/200ug
E-CK-A121 Annexin V-Cyanine5 Reagent 650/670 nm 50T/100T/200T/500T
E-CK-A123 Annexin V-PE/Cyanine5 Reagent 495;565;655/670 nm 50T/100T/200T/500T
E-CK-A125 Annexin V-PE/Cyanine5.5 Reagent 495;565;675/690 nm 50T/100T/200T/500T
E-CK-A127 Annexin V-PE/Cyanine7 Reagent 495;565;755/775 nm 50T/100T/200T/500T
E-CK-A129 Annexin V-APC/Cyanine7 Reagent 650;760/780 nm 50T/100T/200T/500T
E-CK-A131 Annexin V-PE/TR Reagent 495;565/620 nm 50T/100T/200T/500T
E-CK-A133 Annexin V-EV450 Reagent 410/450 nm 50T/100T/200T/500T
E-CK-A133U Annexin V-EV450 Azide-Free Lyophilized Powder 25ug/50ug/100ug/200ug
E-CK-A135 Annexin V-EV500 Reagent 425/500 nm 50T/100T/200T/500T
E-CK-A135U Annexin V-EV500 Azide-Free Lyophilized Powder 25ug/50ug/100ug/200ug
E-CK-A137 Annexin V-AF488 Reagent 495/520 nm 50T/100T/200T/500T
E-CK-A137U Annexin V-AF488 Azide-Free Lyophilized Powder 25ug/50ug/100ug/200ug
 
表五:Annexin V 搭配其他染劑訂購資訊
貨號 產品名稱 螢光 Ex/Em 包裝
30017 Apoptosis & Necrosis Quantification Kit Apoptosis: FITC Annexin V: 498/517 nm; Necrosis: EthD-III: 279, 532/625 nm 50 assays
30065 Apoptosis and Necrosis Quantitation Kit Plus Apoptosis: CF®488A Annexin V: 490/515 nm; Necrosis: EthD-III: 279, 532/625 nm 50 assays
30018 Apoptotic, Necrotic & Healthy Cells Quantification Kit Apoptosis: FITC Annexin V: 498/517 nm; Necrosis: EthD-III: 279, 532/625 nm; Total cells: Hoechst 33342: 352/462 nm 50 assays
30066 Apoptotic, Necrotic & Healthy Cells Quantitation Kit Plus Apoptosis: CF®488A Annexin V: 490/515 nm; Necrosis: EthD-III: 279, 532/625 nm; Total cells: Hoechst 33342: 352/462 nm 50 assays
30030 Dual Apoptosis Assay with NucView® 488 caspase-3 Substrate and Texas Red-Annexin V NucView® 488: 500/530 nm; Texas Red® Annexin V: 594/615 nm 50 assays
30067 Dual Apoptosis Assay with NucView® 488 caspase-3 Substrate and CF®594-Annexin V NucView® 488: 500/530 nm; CF®594 Annexin V: 593/614 nm 50 assays
30073 Dual Apoptosis Assay with NucView® 488 caspase-3 Substrate and CF®640-Annexin V NucView® 488: 500/530 nm; CF®640R Annexin V: 642/662 nm 50 assays
 
表六:7-AAD 和 Propidium Iodide (PI) 訂購資訊
貨號 產品名稱 螢光 Ex/Em 包裝
40037 7-AAD 546/647 nm 1 mg
40084 7-AAD Solution 1 mL (1 mg/mL)
40016 Propidium Iodide 530/622 nm 100 mg
40017 Propidium Iodide (in Water) 10 mL (1 mg/mL in Water)
40048 Propidium Iodide (in Buffer) 2 mL (50 ug/ml in Buffer)