Protein Ultrastructural Localization

Protein ultrastructural localization is the determination of where a protein resides within cells or tissues at nanometer-scale resolution, linking molecular identity to cellular architecture. It commonly uses fixation and thin sectioning followed by antibody-based immunolabeling, in which antibodies bind the target protein and electron-dense tags such as gold particles mark its position for electron microscopy. By revealing whether proteins occupy membranes, organelles, junctions, or extracellular structures, the method helps connect protein distribution with function, trafficking, and disease-related structural changes. It supports cell biology, developmental studies, pathology, and validation of protein targeting in complex biological samples.

Protein Ultrastructural Localization - Related Videos

Research

JoVE Journal - Biology

The CryoAPEX Method for Electron Microscopy Analysis of Membrane Protein Localization Within Ultrastructurally-Preserved Cells

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Cited by 5 •

2020

This protocol describes the cryoAPEX method, in which an APEX2-tagged membrane protein can be localized by transmission electron microscopy within optimally-preserved cell ultrastructure.

Direct Synthesis of EM-Visible Gold Nanoparticles in Cells for Protein Localization Analysis with Well-Preserved Ultrastructure

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Cited by 2 •

2023

The present protocol describes a clonable electron microscopy labeling technology for detecting metallothionein-tagged proteins in cells using a novel autonucleation suppression mechanism-based gold nanoparticle synthesis technique.

Research

JoVE Journal - Biology
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Ultrastructural Localization of Endogenous LC3 by On-Section Correlative Light-Electron Microscopy

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Cited by 6 •

2023

Here, we present a protocol for optimized on-section correlative light-electron microscopy based on endogenous, fluorescent labeling as a tool to investigate the localization of rare proteins in relation to cellular ultrastructure. The power of this approach is demonstrated by ultrastructural localization of endogenous LC3 in starved cells without Bafilomycin treatment.

Identification of Plasmodesmal Localization Sequences in Proteins In Planta

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Cited by 6 •

2017

Plant intercellular connections, the plasmodesmata (Pd), play central roles in plant physiology and plant-virus interactions. Critical to Pd transport are sorting signals that direct proteins to Pd. However, our knowledge about these sequences is still in its infancy. We describe a strategy to identify Pd localization signals in Pd-targeted proteins.

Research

JoVE Journal - Biology
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Nano-fEM: Protein Localization Using Photo-activated Localization Microscopy and Electron Microscopy

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Cited by 14 •

2012

We describe a method to localize fluorescently tagged proteins in electron micrographs. Fluorescence is first localized using photo-activated localization microscopy on ultrathin sections. These images are then aligned to electron micrographs of the same section.

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