冷光式細胞 ATP 含量檢測試劑盒

Luminescent ATP Detection Assay Kit

貨號

ab113849

包裝

300 Test

廠牌

Abcam

產品簡介

Abcam Luminescent ATP Detection Assay Kit 是一款透過冷光反應定量分析細胞內 ATP (adenosine triphosphate) 含量的檢測套組，可應用於細胞存活率 (cell viability)、細胞毒性 (cytotoxicity) 及能量代謝相關研究。套組利用冷光酵素-發光素反應 (luciferase-luciferin reaction) 產生與細胞內 ATP 含量成正比的冷光訊號。搭配可偵測冷光訊號的微量盤分析儀使用（如 INNO-M、INNO-S 或 INNO-H）即可完成定量分析，整體反應僅需兩次試劑添加與混勻步驟，30 分鐘內即可完成檢測。

Functional Studies - Luminescent ATP Detection Assay Kit (AB113849) ab113849 ATP detection kit cytotoxicity data. 25000 HepG2 cells were seeded into each well, allowed to adhere and treated for 4 hours with 25µM rotenone and vehicle control (DMSO) in glucose based complete media. After treatment, cells were lysed, exposed to the ATP substrate solution and signal was measured on a luminescent counter. Mean and standard deviation is plotted for 3 replicates from each condition. Rotenone induces cytotoxicity in HepG2 cells.
Functional Studies - Luminescent ATP Detection Assay Kit (ab113849). ab113849 ATP detection kit cytotoxicity data. 25000 HepG2 cells were seeded into each well, allowed to adhere and treated for 4 hours with 25µM rotenone and vehicle control (DMSO) in glucose based complete media. After treatment, cells were lysed, exposed to the ATP substrate solution and signal was measured on a luminescent counter. Mean and standard deviation is plotted for 3 replicates from each condition. Rotenone induces cytotoxicity in HepG2 cells.
Functional Studies (Act) - Luminescent ATP Detection Assay Kit (AB113849) Simultaneous quantification of mitochondrial respiration and glycolytic flux. Cellular Energy Flux for HepG2 cells (seeded at 65,000 per well), treated with a combination of drug compounds modulating the ETC (Antimycin A [1 μM] and FCCP [2.5 μM]), shown as a percentage relative to untreated control cells. Comparative measurements were taken with Extracellular Oxygen Consumption Assay (ab197243) (white column) and Glycolysis Assay [Extracellular acidification] (ab197244) (black column) show the shift between mitochondrial respiration and glycolysis and the cellular control of energy (ATP; measured 1h post-treatment using Luminescent ATP Detection Assay kit (ab113849) (striped column)).
Functional Studies (Act) - Luminescent ATP Detection Assay Kit (ab113849). Simultaneous quantification of mitochondrial respiration and glycolytic flux. Cellular Energy Flux for HepG2 cells (seeded at 65,000 per well), treated with a combination of drug compounds modulating the ETC (Antimycin A [1 μM] and FCCP [2.5 μM]), shown as a percentage relative to untreated control cells. Comparative measurements were taken with Extracellular Oxygen Consumption Assay (ab197243) (white column) and Glycolysis Assay [Extracellular acidification] (ab197244) (black column) show the shift between mitochondrial respiration and glycolysis and the cellular control of energy (ATP; measured 1h post-treatment using Luminescent ATP Detection Assay kit (ab113849) (striped column)).

主要特色

  • 靈敏度高（可達 1 pM），且線性偵測範圍廣（0.001–10 µM ATP）。
  • 操作簡便快速，僅需兩次試劑添加與混勻步驟，30 分鐘以內即可完成。
  • 細胞裂解步驟可使 ATP 水解酶 (ATPases) 永久失活，確保後續測得的冷光訊號能真實反映樣本中內源性 ATP 含量。
  • 適用樣本類型廣泛，涵蓋懸浮細胞與貼附型細胞，並已有大量研究文獻支持其應用於多種細胞類型。

實驗注意事項

  • ATP 存在於細胞和微生物。為防止背景干擾，建議進行實驗前，先使用 10% 漂白水清潔實驗桌面和 pipettes。實驗手套則建議預先以 70% 酒精清潔，或頻繁更換手套。使用清潔無菌的 tips 和容器，例如不含 ATP 和核酸酶的耗材。操作與孵育反應期間，請勿將試劑或微量多孔盤敞開。
  • 本試劑盒需搭配具備冷光訊號偵測功能的微量盤分析儀和不透明白色 96 孔盤使用，以獲得最佳檢測靈敏度。
  • 建議於光線較暗的環境下進行操作，避免陽光直射或強光照射。強烈光源可能造成孔盤材質產生磷光 (phosphorescence)，進而造成高背景值，干擾訊號判讀。
  • 實驗開始前，應確保所有試劑均已回溫至室溫，避免使用冰冷試劑進行實驗。
  • Substrate 回溶後穩定度會下降，於 4°C 保存一週後，活性可能減少至僅有原本的 70%；因此建議回溶後，分裝避光保存於 -20°C。

科學背景

ATP (adenosine triphosphate) 是細胞內最主要的能量來源，幾乎所有細胞的代謝活動都仰賴 ATP 提供能量，因此 ATP 含量常作為評估細胞存活狀態與能量代謝活性的直接指標。本套組的檢測原理是利用冷光酵素 (luciferase) 催化發光素 (luciferin) 進行氧化反應，此反應需要 ATP 參與，最終釋放出可偵測的冷光訊號，訊號強度與細胞內的 ATP 含量成正比。相較於 MTT、CCK-8 等透過細胞脫氫酶 (dehydrogenase) 將四唑鹽還原為呈色產物的間接性代謝活性檢測方法，冷光式 ATP 檢測是直接定量細胞內的 ATP 含量，能更直接反映細胞的能量代謝狀態，因此除了細胞存活率與細胞毒性評估外，也廣泛應用於粒線體呼吸作用 (mitochondrial respiration)、糖解作用 (glycolytic flux) 等能量代謝相關研究領域。

套組內容物

ItemQuantityStorage temperature (before prep)Storage temperature (after prep)
Detergent20 mL4ºC4ºC
Substrate Buffer20 mL4ºC4ºC
Substrate (lyophilized)3 vials4ºC-20ºC
ATP standard (lyophilized)1 vial4ºC-20ºC

產品說明書

ab113849 Luminescent ATP Detection Assay Kit
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