Display options
Cite
Rahbar Saadat Y, Saeidi N, Zununi Vahed S, et al. An update to DNA ladder assay for apoptosis detection. Bioimpacts. 2015;5(1):25-8doi: 10.15171/bi.2015.01.
Rahbar Saadat, Y., Saeidi, N., Zununi Vahed, S., Barzegari, A., & Barar, J. (2015). An update to DNA ladder assay for apoptosis detection. BioImpacts : BI, 5(1), 25-8. https://doi.org/10.15171/bi.2015.01
Rahbar Saadat, Yalda, et al. "An update to DNA ladder assay for apoptosis detection." BioImpacts : BI vol. 5,1 (2015): 25-8. doi: https://doi.org/10.15171/bi.2015.01
Rahbar Saadat Y, Saeidi N, Zununi Vahed S, Barzegari A, Barar J. An update to DNA ladder assay for apoptosis detection. Bioimpacts. 2015;5(1):25-8. doi: 10.15171/bi.2015.01. Epub 2015 Feb 21. PMID: 25901294; PMCID: PMC4401164.
Copy Download .nbib Format: NLM
Share it on

Bioimpacts. 2015;5(1):25-8. doi: 10.15171/bi.2015.01. Epub 2015 Feb 21.

An update to DNA ladder assay for apoptosis detection.

BioImpacts : BI

Yalda Rahbar Saadat, Nazli Saeidi, Sepideh Zununi Vahed, Abolfazl Barzegari, Jaleh Barar

Affiliations

  1. Research Center for Pharmaceutical Nanotechnology, Faculty of Pharmacy, Tabriz University of Medical Sciences, Tabriz, Iran.
  2. Student Research Committee, School of Advanced Biomedical Sciences, Tabriz University of Medical Science, Tabriz, Iran.

PMID: 25901294 PMCID: PMC4401164 DOI: 10.15171/bi.2015.01

Abstract

INTRODUCTION: A growing interest in apoptosis, programmed cell death, in the last years is observed and leads to better understanding of molecular mechanisms during cell-cell signaling, cell-environment interaction and screening of drugs. This in turn results in emerging of new assays and development of more accurate kits for fast and early detection of apoptosis. However, their sensitivity and reliability have often been scrutinized. Here we introduce a rapid and improved method of DNA ladder apoptosis assay for evaluating apoptosis in mammalian cells.

METHODS: NIH-3T3 cell line was used in this study. After treatment of cells with apoptotic agent, 500 μM H2O2 at 48 hours, DNA was extracted. Then an update protocol of DNA ladder assay was applied for detection of apoptosis. Flow cytometry and DAPI staining were performed to verify apoptosis.

RESULTS: Primary and late apoptosis in the H2O2-treated cells was determined by flow cytometry analysis. DAPI Staining used to show DNA damage and DNA ladder assay using 1.5% gel electrophoresis showed fragmentation in the DNA of treated cells.

CONCLUSION: In this research we aimed to improve DNA ladder assay to the high quality detection of apoptosis in mammalian cells. In our strategy, employing a practical DNA extraction protocol, DNA ladder assay could be applied as an easy/fast method for apoptosis detection. This improved method is able to detect apoptosis in a cost effective/timely manner without need for commercial kits and special equipment.

Keywords: Apoptosis; Cancer; DNA extraction; DNA ladder assay; Signaling pathway

References

  1. Biotechniques. 1997 Sep;23(3):525-31 - PubMed
  2. Cancer Treat Rev. 2008 Dec;34(8):737-49 - PubMed
  3. Methods Mol Biol. 2004;263:141-60 - PubMed
  4. Cytotechnology. 2012 Jan;64(1):9-14 - PubMed
  5. Blood. 1994 Sep 1;84(5):1415-20 - PubMed
  6. J Exp Clin Cancer Res. 2011 Sep 26;30:87 - PubMed
  7. Cold Spring Harb Protoc. 2011 Jan 01;2011(1):pdb.prot5556 - PubMed
  8. Nucleic Acids Res. 1996 Mar 1;24(5):992-3 - PubMed

Publication Types