TY - JOUR
T1 - Non-enzymatic in vitro DNA labeling and label immunoquantification
AU - Clarke, David Higginson
AU - Pérez-Bello, Dannelys
AU - Riverón-Rojas, Ana María
AU - Jiménez-Alemán, Guillermo
AU - Acosta, Mirta Zayas
AU - Perera, Rafaela Pérez
AU - Carrazana, Marquiza Sablón
AU - Miranda, Nelson Díaz
AU - Rodríguez-Tanty, Chryslaine
AU - Calzado, Esteban Gutiérrez
PY - 2005
Y1 - 2005
N2 - In the present work, a label immunoquantification procedure was developed in order to determine the number of markers introduced into DNA. A non-enzymatic, in vitro labeling method for introducing the p-bromobenzoyl radical (label), through transamination and acylation reactions of the cytidine nucleotides in calf thymus DNA, was carried out. Three spacer arms with different lengths were used for separating the label from the nucleotide and three labeled DNA were obtained. Anti-p-bromobenzoyl chicken IgY polyclonal antibodies were obtained. The antibodies detected the label, into three-labeled DNA, with different sensitivities, in relation to spacer arm length used. About 3-11 labels per 4 × 106 bases into thermally denatured DNA were immunoquantified.
AB - In the present work, a label immunoquantification procedure was developed in order to determine the number of markers introduced into DNA. A non-enzymatic, in vitro labeling method for introducing the p-bromobenzoyl radical (label), through transamination and acylation reactions of the cytidine nucleotides in calf thymus DNA, was carried out. Three spacer arms with different lengths were used for separating the label from the nucleotide and three labeled DNA were obtained. Anti-p-bromobenzoyl chicken IgY polyclonal antibodies were obtained. The antibodies detected the label, into three-labeled DNA, with different sensitivities, in relation to spacer arm length used. About 3-11 labels per 4 × 106 bases into thermally denatured DNA were immunoquantified.
KW - Bisulfite-catalyzed transamination reaction
KW - Chicken IgY polyclonal antibodies
KW - In vitro non-radioactive DNA labeling
KW - Label immunoquantification
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U2 - 10.1081/PB-200041428
DO - 10.1081/PB-200041428
M3 - Article
C2 - 15704493
AN - SCOPUS:19944430699
SN - 1082-6068
VL - 35
SP - 1
EP - 15
JO - Preparative Biochemistry and Biotechnology
JF - Preparative Biochemistry and Biotechnology
IS - 1
ER -