H2a.f/z-egfp
H2a.f/z-egfp - Related Videos
11:43
Easy Measurement of Diffusion Coefficients of EGFP-tagged Plasma Membrane Proteins Using k-Space Image Correlation Spectroscopy
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Cited by 35 •
2014
This paper provides a step by step guide to the fluctuation analysis technique k-Space Image Correlation Spectroscopy (kICS) for measuring diffusion coefficients of fluorescently labeled plasma membrane proteins in live mammalian cells.
10:03
Development of Leishmania Species Strains with Constitutive Expression of eGFP
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2023
Here, we describe the methodology used for generating L. panamensis and L. donovani strains expressing the gene for eGFP as a stable integrated transgene using the pLEXSY system. Transfected parasites were cloned by limiting dilution, and clones with the highest fluorescence intensity in both species were selected for further use in drug screening assays.
10:20
Studying Membrane Protein Trafficking in Drosophila Photoreceptor Cells Using eGFP-Tagged Proteins
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Cited by 1 •
2022
Here, non-invasive methods are described for localization of photoreceptor membrane proteins and assessment of retinal degeneration in the Drosophila compound eye using eGFP fluorescence.
10:54
Reliable Identification of Living Dopaminergic Neurons in Midbrain Cultures Using RNA Sequencing and TH-promoter-driven eGFP Expression
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Cited by 9 •
2017
In Parkinson's Disease (PD), Substantia Nigra (SNc) dopaminergic neurons degenerate, leading to motor dysfunction. Here we report a protocol for culturing ventral midbrain neurons from a mouse expressing eGFP driven by a Tyrosine Hydroxylase (TH) promoter sequence, harvesting individual fluorescent neurons from the cultures, and measuring their transcriptome using RNA-seq.
08:02
Live-cell Imaging of Endocytic Transport using Functionalized Nanobodies in Cultured Cells
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Cited by 2 •
2025
Endocytic and retrograde transport of proteins from the plasma membrane to the trans-Golgi network is essential to maintaining membrane homeostasis and regulating signaling. Here, we describe a method to image and quantify endocytic transport of transmembrane cargo proteins by live-cell microscopy using derivatized anti-GFP nanobodies in HeLa cells.
FAQs
The H2A.F/Z portion directs the fusion protein into nucleosomes, while EGFP supplies the detectable fluorescent signal. Because nucleosomes are associated with chromatin, changes in the position or arrangement of labeled nuclei can be followed optically. This makes the reporter useful for observing nuclear behavior as cells move, divide, or reorganize during development.
Nucleosome incorporation anchors EGFP fluorescence to nuclear chromatin rather than leaving the signal independent of nuclear structure. As a result, fluorescence provides a visual reference for the location and behavior of nuclei within living cells. In developmental studies, that reference helps connect cellular movements and tissue rearrangements with the progression of embryonic or tissue development.
H2a.f/z-egfp supports observation of nuclear and cellular dynamics in living material, whereas fixed samples provide information from selected preserved time points. Live visualization can therefore follow how nuclear positions, divisions, and tissue organization change over time. This temporal perspective complements fixed-sample analysis by linking successive developmental events within the same ongoing process.
The reporter is first expressed so that the H2A.F/Z-EGFP fusion can become associated with nucleosomes. Researchers then image the fluorescent nuclei in living embryos or developing tissues while development proceeds. The resulting observations can be aligned with changes in nuclear position, cell division, tissue organization, and other visible cellular dynamics.
Its nuclear fluorescence can be used to follow nuclear movements, patterns of cell division, tissue organization, and broader cellular dynamics. These observations are especially valuable when developmental structures change continuously. Tracking the labeled nuclei provides spatial and temporal context, allowing researchers to relate where nuclei are located and how they behave to the progression of development.
By marking nuclei throughout living embryos or developing tissues, H2a.f/z-egfp helps researchers examine how cells are positioned and rearranged as tissues form. Nuclear locations provide landmarks for interpreting tissue-level changes, while repeated imaging reveals their progression over time. This connects chromatin-associated fluorescence with the spatial organization and dynamic remodeling of developing tissues.