title: Flow Cytometry Method Analysis of Apoptosis: No Significant Difference Between EDTA and EDTA-free Trypsin Treatment Procedure.
abstract: 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.
authors: Department of Pathophysiology, Institute of Pathology and Pathophysiology, College of Basic Medicine, China Medical University, Shenyang, Liaoning 110001, People's Republic of China xyxu@mail.cmu.edu.cn.
authors: Xiao-yan
authors: XY
authors: Xu
authors: Maternity and Infant Hospital Shenyang, Liaoning 110001, People's Republic of China.
authors: Xiao-cui
authors: XC
authors: Nie
authors: Department of Pathophysiology, Institute of Pathology and Pathophysiology, College of Basic Medicine, China Medical University, Shenyang, Liaoning 110001, People's Republic of China.
authors: Hai-ying
authors: HY
authors: Ma
authors: Department of Pancreatic and Breast Surgery, Shengjing Hospital of China Medical University, Shenyang, Liaoning 110001, People's Republic of China.
authors: Guo-qing
authors: GQ
authors: Song
authors: Department of Pathophysiology, Institute of Pathology and Pathophysiology, College of Basic Medicine, China Medical University, Shenyang, Liaoning 110001, People's Republic of China.
authors: Xiao-tong
authors: XT
authors: Zhang
authors: Department of Pathophysiology, Institute of Pathology and Pathophysiology, College of Basic Medicine, China Medical University, Shenyang, Liaoning 110001, People's Republic of China.
authors: Yu-nan
authors: YN
authors: Jin
authors: Department of Pathophysiology, Institute of Pathology and Pathophysiology, College of Basic Medicine, China Medical University, Shenyang, Liaoning 110001, People's Republic of China.
authors: Yan-qiu
authors: YQ
authors: Yu
mesh_terms: D017209:Apoptosis; D001681:Biological Assay; D018929:Cell Culture Techniques; D045744:Cell Line, Tumor; D002614:Chelating Agents / Q000737:chemistry*; D004492:Edetic Acid / Q000737:chemistry*; D005434:Flow Cytometry; D006801:Humans; D014357:Trypsin / Q000737:chemistry*; D051300:Wiskott-Aldrich Syndrome Protein Family / Q000378:metabolism
chemical_list: D002614:Chelating Agents; C120152:WASF2 protein, human; D051300:Wiskott-Aldrich Syndrome Protein Family; D004492:Edetic Acid; D014357:Trypsin
keywords: Apoptosis; EDTA; Flow cytometry; Trypsin; WAVE2.