Double-activated Adomet Analogues

Double-activated AdoMet analogues are synthetic versions of S-adenosylmethionine designed to transfer chemical groups to biological molecules while retaining a second reactive handle for downstream modification. In a methyltransferase-catalyzed reaction, the enzyme recognizes the analogue and cleaves its sulfonium center, transferring the engineered substituent to a nucleophilic site on DNA, RNA, or a protein; the additional activated group can then participate in a selective chemical coupling reaction. These reagents support bioorthogonal labeling, methyltransferase substrate profiling, and mapping of biomolecular interactions, providing researchers with tools to study epigenetic regulation and other AdoMet-dependent processes in complex biological systems.

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JoVE Journal - Biology
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Immunofluorescent Detection of Two Thymidine Analogues (CldU and IdU) in Primary Tissue

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Cited by 48 •

2010

We have derived a strategy to detect sequential incorporation of thymidine analogues (CldU and IdU) into tissues of adult mice to quantify two successive rounds of cell division. This strategy is useful to detect cell turnover of long-lived tissues, oncogenic transformation, or transit-amplifying cells.

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JoVE Journal - Chemistry

Investigations on the Ga(III) Complex of EOB-DTPA and Its 68Ga Radiolabeled Analogue

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Cited by 2 •

2016

A procedure for the isolation of EOB-DTPA and subsequent complexation with natural Ga(III) and 68Ga is presented herein, as well as a thorough analysis of all compounds and investigations on labeling efficiency, in vitro stability and the n-octanol/water distribution coefficient of the radiolabeled complex.

In Vitro Imaging and Quantification of the Drug Targeting Efficiency of Fluorescently Labeled GnRH Analogues

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Cited by 1 •

2017

Selectively labeled fluorescent GnRH-I, -II and -III derivatives are reliable tools for tracking and quantifying their cellular uptake. This manuscript introduces experiments to visualize, quantify and compare the uptake efficiency of these GnRH conjugates in various cell lines.

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JoVE Journal - Biology
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Live-Cell Imaging of Transcriptional Activity at DNA Double-Strand Breaks

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2021

This protocol presents a new reporter gene system and the experimental setup to detect transcription at DNA double-strand breaks with single-molecule sensitivity.

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JoVE Core - Molecular Biology
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Fixing Double-strand Breaks

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2020

The double-stranded structure of DNA has two major advantages. First, it serves as a safe repository of genetic information where one strand serves as the back-up in case the other strand is damaged. Second, the double-helical structure can be wrapped around proteins called histones to form nucleosomes, which can then be tightly wound to form chromosomes. This way, DNA chains up to 2 inches long can be contained within microscopic structures in a cell. A double-stranded break not only damages...

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