Page 13 - 2016 July 專刊-H-0722

【 Cell Death Detection ELISA PLUS 】 ROC-11774425001

偵測原理與標的 Sample preparation
利用 ELISA 方式，偵測凋亡細胞中的 偵測
Histone-complexed DNA fragments 核 ① After incubating cells with an
apoptosis-inducing agent, pellet the cells
酸片段，如果細胞屬於 Necrotic cell， by centrifugation. For the detection of
則收集藥物刺激後的 supernatant 即可 necrosis retain a sample of the DNA
supernatant, which may contain necrotic
測 得 DNA 核 酸 片段。如 果 細胞 屬 於 DNA that leaked through the membrane 的斷裂
Apoptotic cell，在藥物刺激後，需要先 during the incubation.
裂解細胞再收集supernatant，實驗方法 ② Incubate cells with lysis buffer.
如右圖。

③ Pellet the intact nuclei by
適用的實驗分析方法 centrifugation. Take an aliquot of the
--ELISA supernatant (cell lysate) and determine
the amount of apoptotic nucleosomes
present.
適用的偵測檢體
--培養的細胞 (懸浮 or 貼附)
--Supernatant, serum, plasma ELISA assay

產品優點
• 可作定量分析
• 一次可操作多組樣品
• 可以分辨細胞屬於Nercrosis or
Apoptosis
• 沒有物種的限制

Sensitivity of Cell Death Detection ELISAPLUS. Dose-response experiment analyzed by the Cell Death
Different cell concentrations of U937 cells were incubated Detection ELISAPLUS.
with camptothecin (CAM) (2 μg/ml) or without CAM for 4 h at Result: Amounts of cytoplasmic oligonucleosomes (an
37°C. 20 μl of cell culture supernatant and cell lysates were indicator of apoptosis) increase as CAM concentration
analyzed in the ELISA. Substrate reaction time: 10 min. P increases. Cell culture supernatants removed from the cells
Lysate with CAM, P Lysate without CAM, Supernatant with after treatment (but before lysis) gave no signal, indicating
CAM, Supernatant without CAM. that there are no necrotic cells during the
Result: The ELISA can clearly detect apoptosis-related treatment.
nucleosomes in as few as 600 cells.

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