Literature summary for 3.1.11.2 extracted from

Ligasova, A.; Konecny, P.; Frydrych, I.; Koberna, K.
Cell cycle profiling by image and flow cytometry The optimised protocol for the detection of replicational activity using 5-bromo-2'-deoxyuridine, low concentration of hydrochloric acid and exonuclease III (2017), PLoS ONE, 12, e0175880 .

Search for more literature for 3.1.11.2

Application

Application	Comment	Organism
analysis	cell cycle profiling by image and flow cytometry, optimised protocol for the detection of replicational activity using 5-bromo-2'-deoxyuridine, low concentration of hydrochloric acid and exonuclease III, method evaluation and optimization, detailed overview	Homo sapiens

Localization

Localization	Comment	Organism	GeneOntology No.	Textmining
nucleus	-	Homo sapiens	5634	-

Organism

Organism	UniProt	Comment	Textmining
Homo sapiens	Q9NSU2	-	-

Source Tissue

Source Tissue	Comment	Organism	Textmining
A-549 cell	-	Homo sapiens	-
HeLa cell	-	Homo sapiens	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
additional information	detection of replicational activity in cells using 5-bromo-2'-deoxyuridine (BrdU), a low concentration of hydrochloric acid, and exonuclease III, method evaluation and optimization, detailed overview. The approach is based on the introduction of breaks into the DNA by the low concentration of hydrochloric acid followed by the subsequent enzymatic extension of these breaks using exonuclease III. Already 5 to 10 mM HCl is sufficient for the detection of BrdU labelled DNA if exonuclease III is used. The method has only a minimal effect on the tested protein localisations and is applicable both for formaldehyde- and ethanol-fixed cells. The approach partially also preserves the fluorescence of the fluorescent proteins in the HeLa cells expressing fluorescent ubiquitin cell cycle indicator (FUCCI). In the case of the short labelling pulses that disable the use of 5-ethynyl-2'-deoxyuridine because of the low specific signal, the described method provides a bright signal enabling reliable recognition of replicating cells. The optimized protocol is also successfully tested for the detection of trifluridine, the nucleoside used as an antiviral drug and in combination with tipiracil also for the treatment of some types of cancer	Homo sapiens	?	-	?

Synonyms

Synonyms	Comment	Organism
exonuclease III	-	Homo sapiens

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
37	-	assay at	Homo sapiens

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.5	-	assay at	Homo sapiens

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Release 2023.1 (January 2023)