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Antibody against single-stranded DNA useful for detecting apoptotic cells recognizes hexadeoxynucleotides with
Y Kawarada1, N Miura, T Sugiyama
1Department of Biochemistry, Akita University School of Medicine, 1-1-1 Hondo, Akita 010, Japan.
Journal of Biochemistry
|May 30, 1998
Summary
An improved antibody detects apoptotic cells by specifically targeting single-stranded DNA (ssDNA). This antibody recognizes various ssDNA sequences and sizes, proving useful for identifying cellular apoptosis.
Area of Science:
- Immunology
- Molecular Biology
- Cell Biology
Background:
- Previous research established that antibodies against single-stranded DNA (ssDNA) can detect apoptotic cells.
- Further development is needed to refine antibody production and characterize its antigenic determinants for practical application.
Purpose of the Study:
- To develop an improved method for producing an antibody against single-stranded DNA (ssDNA).
- To investigate the antigenic determinants of this ssDNA antibody for enhanced utility in detecting apoptotic cells.
Main Methods:
- Hyperimmunization of rabbits with DNA-methylated bovine serum albumin complexes to generate IgG antibodies.
- Sandwich ELISA and synthetic polymer assays to determine antibody specificity and binding characteristics.
- Gel electrophoresis retardation assays and Western blot analysis to define minimal binding size and recognize apoptotic DNA fragments.
Main Results:
- An IgG antibody specific to single-stranded DNA (ssDNA) was successfully produced.
- The antibody recognizes ssDNA with diverse base sequences and a minimum length of a hexadeoxynucleotide.
- The antibody detected DNA ladders and oligonucleosomes in apoptotic rat liver cells.
Conclusions:
- The developed antibody is a valuable and specific tool for detecting apoptotic cells.
- Characterization of antigenic determinants enhances the antibody's utility in apoptosis research.