Phospho-stat Detection
Phospho-stat Detection - Related Videos
07:12
Time-resolved Förster Resonance Energy Transfer Assays for Measurement of Endogenous Phosphorylated STAT Proteins in Human Cells
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Cited by 4 •
2021
Time-resolved Förster resonance energy transfer cell-based assay protocols are described for the simple, specific, sensitive, and robust quantification of endogenous phosphorylated signal transducer and activator of transcription (STAT) 1/3/4/5/6 proteins in cell lysates in a 384-well format.
03:40
Evaluation of Stromal Cytokine-Induced Intracellular Protein Phosphorylation Using Phospho-Flow Cytometry
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2025
This video demonstrates the evaluation of stromal cytokine-induced intracellular phosphorylation of proteins in leukemia cells. These intracellular phosphorylated proteins are quantified using the phospho-flow cytometry technique.
01:20
The JAK-STAT Signaling Pathway
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2023
Several cytokine receptors have tightly bound Janus kinase or JAK proteins attached at their cytosolic tail. Small signaling molecules such as cytokines, growth hormones, or prolactins bind to the cytokine receptors and initiate their dimerization. The dimerization brings the cytosolic JAKs together that trans-phosphorylate and activates each other. The activated JAKs now phosphorylate cytosolic tails of the cytokine receptors, which serve as binding sites for adaptor proteins such as SH2...
08:42
Immunostaining Phospho-epitopes in Ciliated Organs of Whole Mount Zebrafish Embryos
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Cited by 3 •
2016
Techniques are described to immunostain phospho-epitopes in whole zebrafish embryos and then conduct two-color fluorescent confocal localization in cellular structures as small as primary cilia. The techniques for fixing and imaging can define the location and kinetics of the appearance or activation of specific proteins.
12:45
Detection and Quantification of Tunneling Nanotubes Using 3D Volume View Images
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Cited by 10 •
2022
Tunneling nanotubes (TNTs) are primarily open-ended F-actin membrane nanotubes that connect neighboring cells, facilitating intercellular communication. The notable characteristic that distinguishes TNTs from other cell protrusions is the hovering nature of the nanotubes between cells. Here, we characterize TNTs by constructing a 3D volume view of confocal z-stack images.
FAQs
Phosphorylation connects receptor-associated kinase activity with STAT-dependent gene regulation. After cytokine, growth factor, or other stimulation, JAK-mediated modification supports STAT dimerization and nuclear entry, allowing the proteins to influence transcription. Measuring the phosphorylated form therefore links an upstream biochemical event to the signaling state that precedes changes in gene expression.
The phosphorylated STAT species provides a direct biochemical readout of signaling activity rather than relying only on later gene-expression changes. Its measurement can reveal whether a stimulus has engaged the pathway and whether that response differs between treatments. This makes phosphorylation status useful for examining signaling changes in both experimental and disease-related contexts.
Interpretation depends on the relationship among the stimulus, receptor-associated kinases, STAT proteins, and their phosphorylation state. Cytokines and growth factors can initiate signaling, while JAK activity contributes to STAT modification. The detected phospho-STAT signal should therefore be considered in the context of the treatment or stimulus used and the specific STAT-related pathway being investigated.
Phospho-specific antibodies support several measurement formats, including Western blotting, flow cytometry, and immunofluorescence. Western blotting can assess phosphorylated proteins in biochemical samples, whereas flow cytometry and immunofluorescence enable detection in cell-based formats. The appropriate choice depends on whether the experiment emphasizes biochemical measurement, cellular analysis, or visualization of signaling-related staining.
A typical workflow links a defined cellular stimulus or treatment to antibody-based measurement of the resulting phosphorylated STAT signal. Researchers select a detection format, apply phospho-specific antibodies, and compare the observed signal across treatments or conditions. This design allows pathway activation or suppression to be evaluated in relation to the experimental stimulus.
This approach is used to characterize pathway activation, compare cellular responses to different treatments, and investigate signaling changes associated with disease. It also supports evaluation of inhibitors directed at JAK-STAT and related biochemical pathways. By measuring a signaling intermediate, researchers can connect treatment effects with changes occurring within the pathway rather than considering only downstream outcomes.