Dutp Labeling
Dutp Labeling - Related Videos
05:32
Terminal Transferase-Mediated dUTP Nick End-Labeling—TUNEL Assay: An In Situ Method to Detect DNA Fragmentation in Apoptotic Cells
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2025
In this video, the TUNEL assay is performed to detect the presence of apoptotic cells in a sample. The study involves using a special DNA polymerase - terminal deoxynucleotidyl transferase - that catalyzes the addition of labeled dUTPs to the 3' terminus of fragmented DNA, which can be observed and identified under a fluorescence microscope.
07:37
Using Terminal Transferase-mediated dUTP Nick End-labelling (TUNEL) and Caspase 3/7 Assays to Measure Epidermal Cell Death in Frogs with Chytridiomycosis
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Cited by 3 •
2018
We quantify epidermal cell death in frogs with chytridiomycosis using two methods. First, we use terminal transferase-mediated dUTP nick end-labelling (TUNEL) in situ histology to determine differences between clinically infected and uninfected animals. Second, we conduct a time series analysis of apoptosis over infection using a caspase 3/7 protein analysis.
09:19
Simultaneous Label-Free Autofluorescence Multi-Harmonic Microscopy
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Cited by 3 •
2025
This protocol presents a step-by-step guide for the Simultaneous Label-free Autofluorescence Multi-harmonic (SLAM) microscopic technique, including details on how to generate the laser light source, prepare a tissue sample, conduct imaging, and analyze the data. SLAM advances nonlinear microscopy by measuring four complementary label-free contrasts to investigate the tissue microenvironment.
08:28
Metabolic Labeling
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2023
Metabolic labeling is used to probe the biochemical transformations and modifications that occur in a cell. This is accomplished by using chemical analogs that mimic the structure of natural biomolecules. Cells utilize analogs in their endogenous biochemical processes, producing compounds that are labeled. The label allows for the incorporation of detection and affinity tags, which can then be used to elucidate metabolic pathways using other biochemical analytical techniques, such as SDS-PAGE...
12:16
Robust 3D DNA FISH Using Directly Labeled Probes
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Cited by 49 •
2013
We describe a robust and versatile protocol for analyzing nuclear architecture by 3D DNA FISH using directly labeled fluorescent probes.
FAQs
DNA polymerases accept dUTP in place of thymidine triphosphate during synthesis, so the uridine-containing nucleotide can be incorporated into the growing DNA strand. This substitution creates a route for attaching biotin, digoxigenin, or fluorescent groups to the product. The resulting labeled strand can then be detected after synthesis, linking nucleotide incorporation to assay readout.
Biotin and digoxigenin provide attached recognition groups that can be detected with binding reagents, whereas fluorescent groups can be observed through imaging. Thus, the tag determines whether the experiment relies on a binding-based detection step or fluorescence-based visualization. This distinction helps match the labeled DNA to the intended assay readout.
In DNA-fragmentation assays, dUTP labeling marks DNA associated with fragmentation so it can be identified in the experimental sample. Detection of the incorporated label provides a way to visualize or quantify the fragmentation-related DNA signal. This makes the method useful when the biological question depends on locating or measuring fragmented DNA rather than simply producing a DNA probe.
A typical workflow first uses DNA synthesis to incorporate dUTP or a tagged analog into the probe product. The labeled DNA is then applied in an assay such as in situ hybridization, where detection relies on binding or imaging reagents. The sequence of incorporation followed by assay-specific detection converts an otherwise difficult-to-see probe into a trackable molecular signal.
It is useful when the PCR product needs to be detected through an incorporated label rather than only by its presence as unlabeled DNA. Because polymerases can substitute dUTP for thymidine triphosphate, the amplification product can carry biotin, digoxigenin, or fluorescent groups. A compatible binding or imaging readout then reveals the product.
In situ hybridization uses labeled DNA probes to reveal where specific DNA targets are located in cells or experimental samples. Other assays can use the same labeling principle for PCR product detection or DNA-fragmentation analysis. Depending on the tag and detection reagent, the outcome may be visualization, localization, or quantification of the target-associated DNA signal.