Protein Isolation and Western Blot Analysis

Cells were lysed in cell lysis buffer for the isolation of whole cell proteins, and protein levels were estimated by using the BioRad BCA assay (Biorad Lab Inc. CA) as previously described [43 (link), 44 (link)]. Equal amounts of protein were separated by SDS-PAGE and transferred to polyvinylidene difluoride (PVDF) membranes. The overexpression of the isoforms in MCF-7SphK1a and MCF-7SphK1b was verified by western blot analysis using anti-Flag m2 mouse F1804-1MG from Sigma-Aldrich.

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Chen H., Haddadi N., Zhu X., Hatoum D., Chen S., Nassif N.T., Lin Y, & McGowan E.M. (2022). Expression Profile of Sphingosine Kinase 1 Isoforms in Human Cancer Tissues and Cells: Importance and Clinical Relevance of the Neglected 1b-Isoform. Journal of Oncology, 2022, 2250407.

Publication 2022

BioRad BCA assay

Assay Buffer Cell isolation Cells Isoforms Membranes Mouse Polyvinylidene difluoride Protein Sds page Western blot analysis

Corresponding Organization : Guangdong Pharmaceutical University

Other organizations : The First Affiliated Hospital, Sun Yat-sen University

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