Flow Cytometry Method Analysis of Apoptosis: No Significant Difference Between EDTA and EDTA-free Trypsin Treatment Procedure.

Xiao-yan Xu, Xiao-cui Nie, Hai-ying Ma, Guo-qing Song, Xiao-tong Zhang, Yu-nan Jin, Yan-qiu Yu

Technology in cancer research & treatment 2015 Apr

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Flow cytometry method (FCM) is a generally accepted tool to analyze apoptosis. Although apoptosis assay kit was applied by many companies, the manufacturers were not consistent with whether using Trypsin with EDTA to collect the adherent cells. In another words, the influence of EDTA on apoptotic ratio is not clear. In this work, we compared the proportion of apoptotic cells with EDTA or EDTA-free Trypsin treatment by FCM. We concluded that Trypsin with or without EDTA has little influence on the proportion of apoptotic cells. In addition, we found that the ratio of necrosis and apoptosis was different in cells collected by scraping. WAVE2 protein was analyzed as a typical example for movement related protein. WAVE2 expression is elevated in the EDTA Trypsin treated group, compared with EDTA-free Trypsin treatment and scrapping group. © The Author(s) 2014.

Citation

Xiao-yan Xu, Xiao-cui Nie, Hai-ying Ma, Guo-qing Song, Xiao-tong Zhang, Yu-nan Jin, Yan-qiu Yu. Flow Cytometry Method Analysis of Apoptosis: No Significant Difference Between EDTA and EDTA-free Trypsin Treatment Procedure. Technology in cancer research & treatment. 2015 Apr;14(2):237-41