Multiplexed Phosphoprotein
Multiplexed Phosphoprotein - Related Videos
10:17
A Mass Spectrometry-Based Approach to Identify Phosphoprotein Phosphatases and their Interactors
0 Views •
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.
13:14
Genetic Barcoding with Fluorescent Proteins for Multiplexed Applications
0 Views •
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.
08:05
Multiplexed Fluorometric ImmunoAssay Testing Methodology and Troubleshooting
0 Views •
Cited by 19 •
2011
Using Luminex Corporation’s xMAP microsphere technology, we have developed the Multiplexed Fluorometric ImmunoAssay (MFIA) for serosurveillance of various laboratory animal species. The MFIA is a suspension microarray where antigen, tissue control or immunoglobulins are covalently linked to color-coded polystyrene microspheres. The MFIA testing method as well as various troubleshooting topics is addressed.
05:22
A Multiplex Serological Assay for the Detection of Antibody Responses to Arboviruses
0 Views •
2025
Multiplex fluorescence immunoassays are becoming a reference for serology testing. Their application, however, is limited by the complexity and time-consuming bead-analyte coupling processes. This report presents a semi-automated, reproducible multiplex microsphere bead coupling protocol that is comparable to the standard method while considerably reducing time to results.
08:43
Highly Multiplexed, Super-resolution Imaging of T Cells Using madSTORM
0 Views •
Cited by 5 •
2017
We demonstrate a method to image multiple molecules within heterogeneous nano-structures at single molecule accuracy using sequential binding and elution of fluorescently labeled antibodies.
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.