Mcherry-caax
Mcherry-caax - Related Videos
12:02
Using Tg(Vtg1:mcherry) Zebrafish Embryos to Test the Estrogenic Effects of Endocrine Disrupting Compounds
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Cited by 1 •
2020
Present here is a detailed protocol for the use of zebrafish embryos Tg(vtg1: mCherry) for the detection of estrogenic effects. The protocol covers the propagation of the fish and treatment of embryos, and emphasizes the detection, documentation, and the evaluation of fluorescent signals induced by endocrine disrupting compounds (EDC).
09:32
Light-mediated Reversible Modulation of the Mitogen-activated Protein Kinase Pathway during Cell Differentiation and Xenopus Embryonic Development
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Cited by 7 •
2017
This protocol describes an optogenetic strategy to modulate mitogen-activated protein kinase (MAPK) activity during cell differentiation and Xenopus embryonic development. This method allows for the reversible activation of the MAPK signaling pathway in mammalian cell culture and in multicellular live organisms, like Xenopus embryos, with high spatial and temporal resolution.
10:10
Observing Mitotic Division and Dynamics in a Live Zebrafish Embryo
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Cited by 8 •
2016
Mitosis is critical to every living organism and defects often lead to cancer and developmental disorders. Using this imaging protocol and zebrafish as a model system, researchers can visualize mitosis in a live vertebrate organism and the multitude of defects that arise when mitotic processes are defective.
08:02
Live-cell Imaging of Endocytic Transport using Functionalized Nanobodies in Cultured Cells
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Cited by 1 •
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.
03:01
Calcium Imaging in Individual Neurons Regulating Egg-Laying Behavior in Caenorhabditis elegans
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2025
This video demonstrates a technique for calcium imaging in individual neurons of behaving Caenorhabditis elegans using genetically encoded fluorescent reporter proteins. It outlines the steps involved in imaging worm movement and measuring reporter fluorescence ratios from neurons to correlate intracellular calcium levels with neuronal activity and behavioral changes during egg-laying.
FAQs
The CAAX sequence acts after translation: enzymes recognize its terminal signal and attach a lipid, typically a farnesyl group. This modification increases association of the fluorescent fusion protein with membranes, placing the reporter on their cytoplasmic face. Consequently, fluorescence follows membrane-associated geometry rather than remaining distributed uniformly throughout the cell.
Because the marker associates with the cytoplasmic face, its signal outlines membrane position from inside the cell. This makes changes in cell boundaries, shape, polarity, and membrane-associated structures easier to relate to cellular organization. The resulting images report membrane geometry and dynamics without implying that the fluorophore occupies the extracellular surface.
Its red fluorescence provides a membrane reference that can be compared with other reporters in the same experiment. Researchers can therefore examine membrane behavior together with gene expression, signaling, or cellular-fate readouts. This multiplexing helps connect where membranes change with which developmental signals or expression patterns occur in the same cells or tissues.
A typical workflow uses a genetically encoded mCherry-CAAX construct, allows the fusion protein to be translated and processed through its CAAX sequence, and then follows red fluorescence during live imaging. Researchers examine the resulting membrane-associated signal over time to evaluate cell boundaries, polarity, membrane dynamics, or tissue-level movements during development.
Researchers choose this marker when developmental events depend on changes in cell shape, polarity, or tissue organization. Live imaging can reveal how individual cell boundaries behave as tissues form and rearrange. The marker is especially useful when the goal is to connect membrane dynamics with morphogenetic movements rather than observe gene expression alone.
Time-resolved fluorescence can show how cells alter their outlines, organize within a tissue, and participate in morphogenetic movements. When paired with reporters of gene expression, signaling, or cellular fate, these observations help relate physical membrane behavior to developmental regulation. The resulting comparison links cellular-scale dynamics with broader tissue organization.