Synthesis and purification of horseradish peroxidase-labeled oligonucleotides for tyramide-based fluorescence in situ hybridization

Authors
van Gijlswijk, RPM van de Corput, MPC Bezrookove, V Wiegant, J Tanke, HJ Raap, AK
Citation
Rpm. Van Gijlswijk et al., Synthesis and purification of horseradish peroxidase-labeled oligonucleotides for tyramide-based fluorescence in situ hybridization, HISTOCHEM C, 113(3), 2000, pp. 175-180
Citations number
12
Categorie Soggetti
Medical Research Diagnosis & Treatment
Journal title
HISTOCHEMISTRY AND CELL BIOLOGY
ISSN journal
09486143 → ACNP
Volume
113
Issue
3
Year of publication
2000
Pages
175 - 180
Database
ISI
SICI code
0948-6143(200003)113:3<175:SAPOHP>2.0.ZU;2-S
Abstract
A method is presented to conjugate horseradish peroxidase (HRP) to oligodeo xynucleotides for fluorescence in situ hybridization assays employing tyram ide signal amplification (TSA). HRP is covalently bound to the oligonucleot ide by thiol ether linkage and purified by high-performance liquid chromato graphy. With TSA detection, a single HRP-labeled oligonucleotide probe is s ufficient for in situ detection of clustered DNA repeat sequences with a de gree of repetition between 20 and 50.