相关实验视频
Updated: May 23, 2025
05:29
Measuring Caspase Activity Using a Fluorometric Assay or Flow Cytometry
Published on: March 24, 2023
4.3K
超灵敏的化学发光探头用于检测卡斯巴-3活性
Rozan Tannous1, Chi Zhang2, Doron Shabat1
1School of Chemistry, Raymond and Beverly Sackler Faculty of Exact Sciences, Tel-Aviv University, Tel Aviv 69978 Israel.
Bioconjugate chemistry
|May 9, 2025
概括
我们开发了一种新的化学发光探针,Ac-DEVD-CL,用于高度敏感地检测caspase-3酶活性. 这种探测器克服了现有的光方法的局限性,使得癌细胞中精确的实时亡成像成为可能.
科学领域:
- 生物化学 生物化学
- 分子生物学分子生物学
- 细胞生物学 细胞生物学
背景情况:
- 卡斯巴-3在细胞亡中至关重要,通过分裂细胞蛋白来执行编程细胞死亡.
- 目前对caspase-3的光探测器由于背景噪音和光散射而具有有限的灵敏度.
- 需要更敏感和选择性的探针来实时监测caspase-3活动.
研究的目的:
- 开发和描述一种新型的化学发光探头Ac-DEVD-CL,用于高度敏感和选择性检测caspase-3活性.
- 克服现有的基于光的探针在亡检测中的局限性.
- 为了验证探头对活细胞中亡的实时成像的实用性.
主要方法:
- 开发一种化学发光探针,Ac-DEVD-CL,基于由caspase-3裂变触发的自焚反应.
- 评估探头的启动反应,对其他蛋白酶的特异性和检测极限.
- 将Ac-DEVD-CL与用于检测caspase-3的商业光探针进行比较.
- 在活的4T1乳腺癌细胞中对探针的验证,这些细胞接受了西斯丁治疗,以图像化.
主要成果:
- 在caspase-3激活时,Ac-DEVD-CL的化学发光信号增加了5000倍.
- 探测器证明了对caspase-3的高特异性,交叉反应性最小.
- 与光探测器相比,Ac-DEVD-CL显示了380倍更高的信号噪声比率和100倍更低的检测极限.
- 在活癌细胞中,Ac-DEVD-CL成功对活癌细胞的亡进行了成像,显示出三级的信号增强.
结论:
- Ac-DEVD-CL是一种高度敏感和选择性的化学发光探针,用于检测caspase-3活性.
- 该探测器在现有光方法上为实时亡成像提供了显著的优势.
- Ac-DEVD-CL是癌症研究,药物发现和治疗监测的宝贵工具.
相关概念视频
03:31Labeling DNA Probes
8.1K
DNA probes are fragments of DNA labeled with a reporter tag to enable their detection or purification. The resulting labeled DNA probes can then hybridize to target nucleic acid sequences through complementary base-pairing, and may be used to recover or identify these regions.
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...
8.1K
01:24Caspases
11.8K
Caspase, a family of cysteine proteases, serve as effectors in apoptosis. The ced3 gene in C.elegans was first identified to be involved in apoptosis. This gene encodes the ced-3 caspase that is similar to the interleukin-1-beta converting enzyme or ICE in mammals. In addition to apoptosis, caspases also function in the inflammatory response. Inflammatory caspases are essential in activating pro-inflammatory cytokines that recruit immune cells and block the replication of pathogens inside...
11.8K