TY - JOUR
T1 - A quantitative assay for fragmented DNA in apoptotic cells
AU - Huang, Peng
AU - Plunkett, William
N1 - Funding Information:
This work was supported in part by Grants CA28596 and CA32839 from the National Cancer Institute, Department of Health and Human Services, and Grant DHP-1 from the American Cancer Society.
PY - 1992/11/15
Y1 - 1992/11/15
N2 - Cleavage of cellular chromatin at internucleosomal sites is a characteristic change of DNA integrity in cells undergoing apoptosis. We have developed an assay for quantitation of internucleosomal DNA fragmentation in apoptotic cells. This technique involves purification of cellular DNA, dephosphorylation of the DNA ends, labeling of DNA with 32P at the 5′-end, gel electrophoresis through agarose, and quantitation of the radioactivity in DNA bands. This assay, which is about 1000- to 2000-fold more sensitive than visualization of DNA bands by ethidium staining, allows the detection of DNA fragments at picogram levels. A method for quantitatively determining the number of fragmented DNA strands is also described. Application of this new assay to evaluate the time course of internucleosomal DNA fragmentation was demonstrated in apoptotic cells induced by an anticancer nucleoside analogue.
AB - Cleavage of cellular chromatin at internucleosomal sites is a characteristic change of DNA integrity in cells undergoing apoptosis. We have developed an assay for quantitation of internucleosomal DNA fragmentation in apoptotic cells. This technique involves purification of cellular DNA, dephosphorylation of the DNA ends, labeling of DNA with 32P at the 5′-end, gel electrophoresis through agarose, and quantitation of the radioactivity in DNA bands. This assay, which is about 1000- to 2000-fold more sensitive than visualization of DNA bands by ethidium staining, allows the detection of DNA fragments at picogram levels. A method for quantitatively determining the number of fragmented DNA strands is also described. Application of this new assay to evaluate the time course of internucleosomal DNA fragmentation was demonstrated in apoptotic cells induced by an anticancer nucleoside analogue.
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U2 - 10.1016/0003-2697(92)90518-C
DO - 10.1016/0003-2697(92)90518-C
M3 - Article
C2 - 1489090
AN - SCOPUS:0026439286
SN - 0003-2697
VL - 207
SP - 163
EP - 167
JO - Analytical Biochemistry
JF - Analytical Biochemistry
IS - 1
ER -