Bioimpacts. 2015;5(1): 25-28.
doi: 10.15171/bi.2015.01
PMID: 25901294
PMCID: PMC4401164
Scopus ID: 84924904679
  Abstract View: 4734
  PDF Download: 2683

Original Research

An update to DNA ladder assay for apoptosis detection

Yalda RahbarSaadat 1, Nazli Saeidi 1, Sepideh Zununi Vahed 2, Abolfazl Barzegari 2*, Jaleh Barar 1 ORCID logo

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
*Corresponding Author: Email: barzegari.abolfazl@gmail.com


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.
First Name
Last Name
Email Address
Security code

Abstract View: 4523

Your browser does not support the canvas element.

PDF Download: 2683

Your browser does not support the canvas element.

Submitted: 20 Feb 2015
ePublished: 15 Aug 2017
EndNote EndNote

(Enw Format - Win & Mac)

BibTeX BibTeX

(Bib Format - Win & Mac)

Bookends Bookends

(Ris Format - Mac only)

EasyBib EasyBib

(Ris Format - Win & Mac)

Medlars Medlars

(Txt Format - Win & Mac)

Mendeley Web Mendeley Web
Mendeley Mendeley

(Ris Format - Win & Mac)

Papers Papers

(Ris Format - Win & Mac)

ProCite ProCite

(Ris Format - Win & Mac)

Reference Manager Reference Manager

(Ris Format - Win only)

Refworks Refworks

(Refworks Format - Win & Mac)

Zotero Zotero

(Ris Format - Firefox Plugin)