通过免疫荧光在乳腺癌细胞中定量 EGFR 内体再循环。
Quantifying EGFR endosomal recycling via immunofluorescence in breast cancer cells.
机构信息
Centre for Cancer Biology, An Alliance of SA Pathology and the University of South Australia, Adelaide, SA, Australia.
Department of Molecular and Biomedical Science, School of Biological Sciences, Faculty of Sciences, University of Adelaide, Adelaide, SA, Australia.
出版信息
STAR Protoc. 2022 Apr 13;3(2):101305. doi: 10.1016/j.xpro.2022.101305. eCollection 2022 Jun 17.
Previously published protocols for quantification of endosomal recycling are limited by the use of radioactive reagents, washing of cells in reducing buffers, or the requirement for large numbers of cells. Here, we describe a protocol for quantification of endosomal recycling using immunofluorescence that is optimized for EGFR in BT-549 breast cancer cells but could be applied to other RTKs and cell lines. Our protocol enables quick assessment of recycling and uses a relatively small number of cells. For complete details on the use and execution of this protocol, please refer to Lonic et al. (2021).
先前发表的用于定量内体再循环的方案受到放射性试剂的使用、在还原缓冲液中洗涤细胞或需要大量细胞的限制。在这里，我们描述了一种使用免疫荧光定量 EGFR 的方案，该方案在 BT-549 乳腺癌细胞中进行了优化，但可应用于其他 RTK 和细胞系。我们的方案能够快速评估再循环，并使用相对较少的细胞。有关此方案使用和执行的完整详细信息，请参阅 Lonic 等人。（2021 年）。
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