Receptor Trafficking Analysis

Receptor trafficking analysis is the study of how cell-surface receptors move through intracellular compartments, a process that regulates signal strength, duration, and cellular responsiveness. Researchers track receptor internalization, sorting, recycling to the plasma membrane, or delivery to lysosomes for degradation using methods such as fluorescence microscopy, labeled ligands, and biochemical assays. In biology, these analyses reveal how cells control communication in response to hormones, neurotransmitters, growth factors, and drugs. They also help explain altered signaling in disease and evaluate therapies designed to change receptor abundance, localization, or activation.

Receptor Trafficking Analysis - Related Videos

Research

JoVE Journal - Biology

Tracking Drug-induced Changes in Receptor Post-internalization Trafficking by Colocalizational Analysis

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Cited by 1 •

2015

Receptor trafficking modulates signaling and cell responsiveness to ligands and is, itself, responsive to cell conditions, including ligand-induced signaling. Here, we describe a powerful and flexible technique for quantitatively assessing drug-induced receptor trafficking using immunolabeling and colocalizational analysis.

Utilizing pHluorin-tagged Receptors to Monitor Subcellular Localization and Trafficking

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Cited by 3 •

2017

Labeling the extracellular domain of a membrane protein with a pH sensitive fluorophore, superecliptic pHluorin (SEP), allows subcellular localization, expression, and trafficking to be determined. Imaging SEP-labeled proteins with total internal reflection fluorescence microscopy (TIRFM) enables the quantification of protein levels in the peripheral ER and plasma membrane.

Analysis of SCAP N-glycosylation and Trafficking in Human Cells

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Cited by 17 •

2016

We describe a modified method for membrane fraction isolation from human cells and sample preparation for the detection of SCAP N-glycosylation and total protein by using western blot. We further introduce a GFP-labeling method to monitor SCAP trafficking using confocal microscopy. This protocol can be used in regular biology laboratories.

Using an α-Bungarotoxin Binding Site Tag to Study GABA A Receptor Membrane Localization and Trafficking

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Cited by 5 •

2014

Here we demonstrate the use of fluorescent Alexa dye coupled to α-bungarotoxin to measure GABA A receptor surface localization and endocytosis in hippocampal neurons. Through the use of constructs bearing a short extracellular tag that binds α-bungarotoxin, analysis of plasma membrane protein endocytic trafficking can be achieved.

Live Cell Imaging and 3D Analysis of Angiotensin Receptor Type 1a Trafficking in Transfected Human Embryonic Kidney Cells Using Confocal Microscopy

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Cited by 2 •

2017

Here we present a protocol to image cells expressing green fluorescent protein-tagged angiotensin type 1a receptors during endocytosis initiated by angiotensin II treatment. This technique includes labeling lysosomes with a second fluorescent marker, and then utilizing software to analyze the co-localization of receptor and lysosome in three dimensions over time.

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