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分析人类肝癌细胞系中能量代谢途径依赖性的协议
Sk Ramiz Islam1, Sebabrata Maity1, Oishee Chakrabarti1
1Biophysics & Structural Genomics Division, Saha Institute of Nuclear Physics, 1/AF Bidhannagar, Kolkata, West Bengal 700 064, India; Homi Bhabha National Institute, BARC Training School Complex, Anushaktinagar, Mumbai, Maharashtra 400 094, India.
STAR protocols
|March 20, 2024
概括
我们开发了一种简单,直接的方法来测量细胞ATP水平,揭示代谢途径的贡献. 这种可访问的协议可以对任何细胞系的细胞能量代谢进行高通量分析.
科学领域:
- 生物化学 生物化学
- 细胞生物学 细胞生物学
- 代谢学 代谢学 代谢学
背景情况:
- 分析细胞能量代谢通常是复杂的,昂贵的和间接的.
- 现有的方法通常依赖于间接测量或复杂的仪器仪表.
- 需要一种直接和可访问的方法来理解代谢途径对ATP生产的贡献.
研究的目的:
- 提出一种新的,高通量协议,用于直接测量ATP水平,以评估细胞能量代谢.
- 为了能够分析代谢途径对细胞腺三酸盐 (ATP) 生产的相对贡献.
- 为干扰研究提供适用于任何细胞系的可访问方法.
主要方法:
- 一个涉及细胞计数,在96孔板中播种和用甲胺治疗的协议.
- 使用特定代谢抑制剂对代谢途径的系统抑制.
- 进行细胞活力测试和腺三酸盐 (ATP) 测试.
- 根据直接的ATP测量计算细胞能量代谢依赖性.
主要成果:
- 展示了测量ATP水平以评估代谢途径贡献的直接方法.
- 成功地应用了该协议来分析各种细胞系中的能量代谢.
- 在灵活扰动研究中验证了该协议的实用性.
结论:
- 本协议提供了一种直接的,高吞吐量和可访问的方法来分析细胞能量代谢.
- 这种方法简化了对代谢途径对ATP生产的贡献的评估.
- 该协议促进了跨多种细胞系的灵活扰动研究.
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