Edu Labeling Assay
Edu Labeling Assay - Related Videos
09:49
Antigen Specific In Vivo Killing Assay using CFSE Labeled Target Cells
0 Views •
Cited by 39 •
2010
Many infections elicit a strong CTL response, but occasionally, the quantity of responding cells does not correlate to control of the pathogen1. One measure of CTL quality is their ability to kill specifically2. CFSE labeling of target cells can be used to investigate this CTL response quality in vivo3,4.
08:18
High-throughput Nitrobenzoxadiazole-labeled Cholesterol Efflux Assay
0 Views •
Cited by 2 •
2019
Measurement of in vitro cholesterol efflux capacity to serum or plasma in macrophage cell models is a promising tool as a biomarker for atherosclerosis. In the present study, we optimize and standardize a fluorescent NBD-cholesterol efflux method and develop a high-throughput analysis using 96-well plates.
03:59
Bromodeoxyuridine Pulse Labelling Assay: A Technique to Measure Cell Proliferation
0 Views •
2023
The video describes the stepwise pulse labeling assay of mammalian cells with bromodeoxyuridine (BrdU) for measuring cell proliferation. The BrdU uptake permits the temporal tracking of cells that were in the synthesis phase at a specific point in time, without requiring the cells to be synchronized.
09:19
Simultaneous Label-Free Autofluorescence Multi-Harmonic Microscopy
0 Views •
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.
09:58
An Optimized Protocol for Electrophoretic Mobility Shift Assay Using Infrared Fluorescent Dye-labeled Oligonucleotides
0 Views •
Cited by 14 •
2016
We describe here an optimized protocol of fluorescent Electrophoretic Mobility Shift Assays (fEMSA) using purified SOX-2 proteins together with infrared fluorescent dye-labeled DNA probes as a case study to tackle an important biological question.
FAQs
During S phase, EdU is incorporated into DNA as replication proceeds, creating a chemical handle for later detection. This design links fluorescence to the DNA-synthesis event itself rather than requiring a primary antibody to recognize a thymidine analog. The assay therefore identifies cells actively engaged in replication during the labeling period.
The post-fixation reaction with a fluorescent azide marks incorporated EdU for detection. Copper-catalyzed click chemistry connects the DNA-synthesis event to a fluorescent readout that can be observed by microscopy or measured by flow cytometry. This separates labeling during replication from signal generation during the later detection step.
Compared with antibody-based thymidine analog assays, EdU labeling removes the need for antibody recognition of the incorporated analog. Detection occurs through a post-fixation reaction with a fluorescent azide, providing a direct workflow compatible with both microscopy and flow cytometry. This supports efficient visualization and quantitative analysis of labeled cells.
Researchers can compare labeled cells across experimental conditions to assess changes in DNA-synthesis activity. Applying the assay to treated and untreated biological samples helps reveal whether a developmental cue, drug, or other condition is associated with altered proliferation. Microscopy or flow cytometry then supplies the corresponding visual or quantitative readout.
Cells first encounter EdU while DNA synthesis is occurring. The biological sample is then fixed, and a fluorescent azide is added for copper-catalyzed click chemistry with the incorporated label. Finally, researchers examine the sample by microscopy or quantify labeled cells with flow cytometry, depending on the experimental goal.
Microscopy allows researchers to visualize which cells in a biological sample contain the EdU-associated fluorescent signal. Flow cytometry provides a quantitative approach for assessing labeled cells across the sample. Using either platform, or selecting between them according to the study design, connects detection of DNA synthesis with analysis of proliferation.