Multiplexed Phosphoprotein
Multiplexed Phosphoprotein - Related Videos
10:17
A Mass Spectrometry-Based Approach to Identify Phosphoprotein Phosphatases and their Interactors
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2022
Here, we present a protocol for the enrichment of endogenous phosphoprotein phosphatases and their interacting proteins from cells and tissues and their identification and quantification by mass spectrometry-based proteomics.
04:21
Multiplex Cyclic Fluorescent Immunohistochemistry
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Cited by 1 •
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Multiplex cyclic immunohistochemistry allows in situ detection of multiple markers simultaneously using repeated antigen-antibody incubation, image scanning, and image alignment and integration. Here, we present the operating protocol for identifying immune cell substrates with this technology in lung cancer and paired brain metastasis samples.
05:25
Simultaneous Detection of Different Antibody Classes in a Multiplexed Serological Test
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2023
A three-channel dual-reporter fluorescence flow analysis system was used to develop a bead-based multiplex immunoassay that simultaneously evaluates serum samples for IgG and IgM elicited against multiple antigens of different Borrelia species that cause Lyme borreliosis in Europe and North America.
07:18
Highly-Multiplexed Tissue Imaging with Raman Dyes
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Cited by 5 •
2022
Electronic pre-resonance stimulated Raman scattering (epr-SRS) imaging of rainbow-like Raman dyes is a new platform for highly multiplexed epitope-based protein imaging. Here, we present a practical guide including antibody preparation, tissue sample staining, SRS microscope assembly, and epr-SRS tissue imaging.
13:14
Genetic Barcoding with Fluorescent Proteins for Multiplexed Applications
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Cited by 4 •
2015
Since the discovery of the green fluorescent protein gene, fluorescent proteins have impacted molecular cell biology. This protocol describes how expression of distinct fluorescent proteins through genetic engineering is used for barcoding individual cells. The procedure enables tracking distinct populations in a cell mixture, which is ideal for multiplexed applications.
FAQs
Its main advantage is parallel pathway coverage. By measuring phosphorylation states across several proteins in one sample, researchers can compare coordinated changes rather than interpreting a single signaling event in isolation. This broader profile can reveal whether an experimental response affects multiple signaling pathways and helps relate molecular changes to larger cellular processes.
Target-specific antibodies and related affinity reagents provide the molecular selectivity needed to distinguish phosphorylated proteins within a complex sample. They capture or detect selected targets simultaneously, making it possible to examine several signaling components in the same analysis. The resulting measurements support comparisons among proteins, pathways, and experimental conditions.
Condition-to-condition comparisons show how signaling changes in response to a defined biological context. Instead of considering phosphorylation measurements as isolated values, researchers can identify patterns associated with stimulation, injury, or disease-related processes. These patterns help connect altered signaling networks with changes in neuronal behavior, survival, or synaptic function.
A multiplexed profile places phosphorylation changes from several proteins into the same analytical context. If multiple targets change together, the pattern can indicate broader pathway-level remodeling; if only one target changes, the result may represent a more restricted response. In neuroscience, this distinction helps interpret kinase-driven effects across neural tissues.
The workflow begins with a sample containing the proteins of interest, followed by simultaneous target-specific capture or detection using antibodies or other affinity reagents. Phosphorylation states are then measured across the selected targets and compared between experimental conditions. This approach produces a multi-target signaling profile rather than a result for only one protein.
It is particularly useful when researchers need to characterize signaling responses in neurons or neural tissues after stimulation, injury, or disease-related change. Measuring several phosphoproteins at once can reveal kinase-driven patterns linked to synaptic function and neuronal survival, while also highlighting signaling components that may warrant consideration as therapeutic targets.