All rights reserved. EdU is a nucleoside analog of thymidine and is incorporated into DNA during active DNA synthesis.1 Detection of EdU is based on a click reaction,1-5 which is a copper (I) catalyzed reaction between an azide and an alkyne.
The use of Invitrogen Alexa Fluor dyes results in a bright, photostable fluorescent signal designed for easy detection of proliferating cells.The Invitrogen Click-iT EdU Colorimetric IHC Detection Kit provides a simple, robust method for analyzing DNA replication in proliferating cells in as little as 90 minutes. This method makes it possible to assess cell proliferation with microscopic detection at cellular level and routine screening. Cells that have incorporated BrdU into their DNA are easily detected using a monoclonal …
The use of Invitrogen Alexa Fluor dyes results in a bright, photostable fluorescent signal designed for easy detection of proliferating cells.The Invitrogen Click-iT EdU Colorimetric IHC Detection Kit provides a simple, robust method for analyzing DNA replication in proliferating cells in as little as 90 minutes. Assays can report either total cell numbers or live cells, or measure DNA synthesis in single cells.Our broad range of Invitrogen cell analysis assays and fluorescent labels can be used individually or together to carry out complex biological investigations, including studies of cell proliferation, cytotoxicity, or drug efficacy, using imaging, microplate, or flow cytometry assay platforms.Measuring the synthesis of new DNA is a precise way to assay cell proliferation in individual cells or in cell populations. DNA polymerase in the replisome synthesizes DNA on resulting ssDNA as a template, resulting in lengthening of DNA ( Fig. Proliferation measurements are typically made based on average DNA content or on cellular metabolism. A10044,C10269,C10214,O10180,A889,A21253,T10182,A6397,T20915,T20922,T20923,T30954 Because cellular DNA content is highly regulated, the CyQUANT assay can be used at multiple time points to calculate the average proliferation rate of a cell population. Traditionally, radiolabeled 3H-thymine is incubated with cells for several hours or overnight. Single-strand binding protein (SSB) is associated with resulting ssDNA before DNA synthesis by DNA polymerase and is also involved in DNA repair and recombination. We next wanted to determine whether EdU can be used to detect DNA synthesis in animals. A new technique that combines the Traditional methods for monitoring nascent nuclear DNA synthesis follow the incorporation of a nucleoside analog of thymidine into DNA.
The regulation of mtDNA replication and its role in mitochondrial biogenesis remains a mystery [1].
The newly proliferated DNA cells will incorporate the radiolabels and can be detected by a scintillation counter after elution.
into adult mice and tissues were harvested and fixed 96 h after injection. This assay eliminates not only the use of radioactivity but also the harsh DNA denaturation step required with the BrdU method (Figure 1). EdU detection is based on a click reaction—a copper(I)-catalyzed reaction between an azide and an alkyne [3]. Changes in mitochondrial DNA (mtDNA) copy number are not only important for normal biological processes, but are also implicated in drug toxicity, aging, and a wide range of diseases including cancer, diabetes, and neurodegeneration.
New DNA synthesis—imaging/high-content analysis Measuring the synthesis of new DNA is a precise way to assay cell proliferation in individual cells or in cell populations. DNA Synthesis Proliferation Assays. Because of the high labeling efficiency and the lack of harsh treatments, the Click-iT EdU Assays provide a superior alternative to traditional proliferation assays.
Colorimetric signal is perfect for tissues with high background fluorescence; the proliferation signal that can be archived for future analysis.The Invitrogen Click-iT Plus EdU Assay Kits provide simple, robust methods for analyzing DNA replication in proliferating cells in as little as 90 minutes. Detection is achieved through a copper-catalyzed “click” reaction that is complete typically within …
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