Quantitative Western Blot Analysis

Western blot was performed according to our previous paper [4 (link)]. In brief, whole cell extracts were prepared by lysing the cells in lysis buffer (KeyGEN biotech, Nanjing, China). Then the equal amounts of total proteins were resolved by SDS-PAGE and the proteins of interest were probed by Western blot. Subsequently the Western blot results were visualized by enhanced chemiluminescence according to manufacturer’s instructions (Millipore, Billerica, MA, USA). The expression of protein was quantified by Image J software.

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Luo W., Chang G., Lin D., Xie H., Sun H., Li Z., Mo S., Wang R., Wang Y, & Zheng Z. (2024). 3,3’-((3,4,5-trifluoropHenyl)methylene)bis(4-hydroxy-2H-chromen-2-one) inhibit lung cancer cell proliferation and migration. PLOS ONE, 19(5), e0303186.

Publication 2024

lysis buffer enhanced chemiluminescence

Corresponding Organization : Guangdong Pharmaceutical University

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Variable analysis

independent variables

Preparation of whole cell extracts by lysing the cells in lysis buffer

dependent variables

Expression of protein of interest
Quantification of protein expression using Image J software

control variables

Equal amounts of total proteins resolved by SDS-PAGE
Visualization of Western blot results by enhanced chemiluminescence according to manufacturer's instructions

controls

No positive or negative controls were explicitly mentioned in the protocol.

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