Brdu Immunohistochemistry
Brdu Immunohistochemistry - Related Videos
08:37
Immunohistochemistry Techniques to Analyze Cellular Proliferation and Neurogenesis in Rats Using the Thymidine Analog BrdU
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Cited by 7 •
2020
This paper presents four of the most common techniques for visualizing BrdU positive cells to measure adult neurogenesis in rats. This work includes instructions for reagent preparation, thymidine analog administration, transcardial perfusion, tissue preparation, peroxidase immunohistochemical reaction, immunofluorescence, signal amplification, counterstaining, microscopy imaging, and cell analysis.
03:24
BrdU Immunofluorescence Staining: A Technique to Identify Cells in Different Phases of Cell Cycle
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2023
This video describes the protocol for BrdU immunofluorescence staining. The technique allows the identification of cells in different phases of cell cycle. Addition of DNA dyes and antibody labeling allows the detailed analysis of the fate of the S phase cells at later times.
09:14
Examination of Proteins Bound to Nascent DNA in Mammalian Cells Using BrdU-ChIP-Slot-Western Technique
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Cited by 4 •
2016
In this protocol, we describe a novel BrdU-ChIP-Slot-Western technique to examine proteins and histone modifications associated with newly synthesized or nascent DNA.
08:29
Drosophila Pupal Abdomen Immunohistochemistry
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Cited by 20 •
2011
Antibody staining of the Drosophila pupae can enhance genetic analyses of adult abdominal developmental genetics. We present our protocol for dissection, fixation and antibody staining of staged Drosophila pupal abdomen.
10:10
Drosophila Larval NMJ Immunohistochemistry
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Cited by 26 •
2009
This protocol demonstrates how to perform immunohistochemistry on dissected Drosophila larva.
FAQs
The S-phase dependence creates a time-linked record of DNA synthesis. Only cells replicating DNA during BrdU exposure can acquire the analog, so labeled nuclei indicate participation in that replication window rather than a general property of all cells. In neuroscience, this timing helps associate labeling with proliferative activity in neural progenitor populations.
DNA denaturation is essential because it makes the incorporated BrdU accessible to the anti-BrdU antibody. Without this tissue-processing step, antibody binding and the resulting signal may not adequately reveal labeled nuclei. Thus, denaturation connects the biochemical event in newly synthesized DNA to the visual readout used for identifying proliferating cells.
Chromogenic and fluorescent signals are alternative readouts for antibody-bound BrdU. The choice determines how labeled nuclei are visualized, while the underlying evidence remains antibody recognition of incorporated analog in denatured DNA. This distinction matters when interpreting images, because the signal format changes the readout without changing the replication event being marked.
A positive nucleus records DNA replication during the BrdU exposure period, but the biological question can extend beyond that initial event. Examination of labeled cells in developing, healthy, or injured nervous tissue can support analyses of whether newly generated cells persist and whether they undergo later differentiation. Thus, labeling supplies a starting point for cell-fate studies.
A practical workflow links labeling, tissue preparation, and antibody detection in sequence. Animals or cells first receive BrdU, tissue is then processed so incorporated DNA becomes accessible through denaturation, and anti-BrdU antibodies reveal labeled nuclei with chromogenic or fluorescent signals. Keeping these stages conceptually distinct helps researchers connect the final image to the original replication event.
In neuroscience, the technique is useful when the question concerns newly generated cells in the nervous system. It can be applied to developing tissue, healthy tissue, or tissue after injury, allowing researchers to identify proliferating neural progenitors and examine adult neurogenesis. The same labeling framework also supports studies of survival or differentiation after cell generation.