Phospho Jacob Detection
Phospho Jacob Detection - Related Videos
10:52
Radiolabeling and Quantification of Cellular Levels of Phosphoinositides by High Performance Liquid Chromatography-coupled Flow Scintillation
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Cited by 21 •
2016
Phosphoinositides are signaling lipids whose relative abundance rapidly changes in response to various stimuli. This article describes a method to measure the abundance of phosphoinositides by metabolically labeling cells with 3H-myo-inositol, followed by extraction and deacylation. Extracted glycero-inositides are then separated by high-performance liquid chromatography and quantified by flow scintillation.
06:23
pH Modulation Assay on Supported Lipid Bilayers to Detect Protein-Phosphoinositide Interactions
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2025
In this video, we demonstrate protein-phosphoinositide interactions in a microfluidic pattern using the pH modulation assay, using pH-sensitive fluorescent dye tagged to phosphatidylethanolamine lipid of a supported lipid bilayer, called a microfluidic platform. The modulations in pH after protein binding to phospholipid cause fluorescence quenching.
01:42
Phosphoinositides and PIPs
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2023
Phosphoinositides are a group of phospholipids containing a glycerol backbone with two fatty acid chains and a phosphate attached to a myoinositol sugar ring. The inositol head group extends into the cytoplasm, where it is modified by adding phosphate groups to form phosphatidylinositol phosphates or PIPs. Different phosphoinositides are synthesized and recruited on the cytosolic face of the plasma membrane. The localization of specific phosphoinositides concentrated in separate membrane...
10:58
PIP-on-a-chip: A Label-free Study of Protein-phosphoinositide Interactions
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Cited by 7 •
2017
Here we present a supported lipid bilayer in the context of a microfluidic platform to study protein-phosphoinositide interactions using a label-free method based on pH modulation.
09:10
Combination of Adhesive-tape-based Sampling and Fluorescence in situ Hybridization for Rapid Detection of Salmonella on Fresh Produce
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Cited by 17 •
2010
This protocol describes a simple adhesive-tape-based approach for sampling of tomato and other fresh produce surfaces, followed by rapid whole cell detection of Salmonella using fluorescence in situ hybridization (FISH).
FAQs
Synaptic stimulation can promote phosphorylation of Jacob, creating a molecular change that connects activity at synapses with downstream nuclear signaling. Phosphorylated Jacob may then be transported to the neuronal nucleus, where its presence provides evidence that synaptic activity has engaged activity-dependent gene-regulatory pathways. This makes the signal useful for studying how neuronal communication produces longer-lasting cellular responses.
Nuclear transport gives phospho-Jacob detection a spatial dimension beyond measuring phosphorylation alone. Finding phosphorylated Jacob in the neuronal nucleus can indicate that activity-dependent signaling has progressed from synaptic regions toward gene regulation. Consequently, cellular distribution may help distinguish where signaling occurs and how neuronal activity is linked to changes associated with plasticity.
Phosphorylated Jacob serves as a readout of signaling associated with activity-dependent plasticity. Its abundance can indicate how much of the phosphorylated form is present, while its distribution can show where that signaling component is located within neurons. Together, these measurements help connect synaptic stimulation with cellular processes relevant to changes in neuronal function.
Immunofluorescence can show the cellular distribution of phosphorylated Jacob, including its relationship to neuronal regions such as the nucleus. Western blotting is suited to measuring its abundance in an assay sample. Thus, the methods address complementary questions: one emphasizes spatial localization, whereas the other emphasizes the amount of the detected phosphorylated protein.
The central approach is to apply antibodies specific to phosphorylated Jacob in an appropriate detection assay. Immunofluorescence is used when cellular localization is important, whereas Western blotting supports measurement of phospho-Jacob abundance. Related assays may also be used, provided they detect the phosphorylated form and support the intended question about signaling or distribution.
Researchers can use phospho-Jacob detection when they need to examine activity-dependent signaling after synaptic stimulation or assess molecular events associated with synaptic plasticity. The approach is also relevant to investigations of learning, memory, neurodevelopment, and neurological disease. Depending on the assay, results can map cellular distribution, estimate abundance, or connect neuronal activity with gene-regulatory signaling.