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7.0K Views
•
09:47 min
February 1st, 2019
DOI :
10.3791/58966-v
Chapters
0:04
Title
1:05
Generation of LN215-YFPQL and LN215-I" Cells by Lentiviral Transduction
3:14
Plating the LN215-YFPQL and LN215-I‐ Cells
4:42
Conducting the I-YFP GJIC Assay
7:37
Results: Analysis of the Iodine-Yellow Fluorescent Protein-Gap Junction Intercellular Communication Assay
8:15
Conclusion
Transcript
差距交界处已被建议为药物目标。碘化物-YFP-GJIC测定与高通量筛选兼容,可发现间隙结调制剂。它被用来在相对较短的时期内测试大量化合物对间隙结活动的影响。
碘化物-YFP-GJIC测定简单、坚固、可重复且价格低廉。这种测定只需要接受方和供体细胞和两个平衡的盐溶液。无需其他试剂,如路西法黄色和钙素 AM。
它还测量单个井中细胞的总间隙结活动,从而导致井间变异性低。为了开始此过程,如文本协议中所述,在补充的 DMEM 中将 LN215 细胞增长到 80% 的汇合。转导
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Summary
在这里, 我们提出了一种新的缝隙结细胞间通信检测的协议, 旨在为药物发现和毒理学评估的间隙结调节化学品的高通量筛选。
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