Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

Protein Dynamics in Living Cells

Protein Dynamics in Living Cells

Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...

Photoluminescence: Applications

Photoluminescence: Applications

Photoluminescence offers a wide range of applications due to its inherent sensitivity and selectivity. This technique allows for both direct and indirect analyses of the analyte. Direct quantitative analysis is possible when the analyte exhibits a favorable quantum yield for fluorescence or phosphorescence. However, an indirect analysis may be feasible if the analyte is not fluorescent or phosphorescent, or if the quantum yield is unfavorable. Indirect methods include reacting the analyte with...

Labeling DNA Probes

Labeling DNA Probes

DNA probes are fragments of DNA labeled with a reporter tag to enable their detection or purification. The resulting labeled DNA probes can then hybridize to target nucleic acid sequences through complementary base-pairing, and may be used to recover or identify these regions.
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...

Western Blotting

Western Blotting

Western blotting is an analytical technique for protein identification. It has various applications in immunology and medicine, including detecting diseases like bovine spongiform encephalopathy, mad cow disease, and human and feline immunodeficiency virus from biological samples.
The technique begins with separating proteins from the sample using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), followed by protein transfer, immunoblotting, and finally, protein detection.

Fluorescence and Phosphorescence: Instrumentation

Fluorescence and Phosphorescence: Instrumentation

Fluorometers and spectrofluorometers are two types of instruments used for measuring molecular fluorescence. These instruments differ in how they select excitation and emission wavelengths and the type of light sources they utilize. Fluorometers use absorption interference filters to choose excitation and emission wavelengths. The excitation source in a fluorometer is typically a low-pressure mercury vapor lamp that emits intense lines distributed throughout the ultraviolet and visible regions.

Phosphorylation

Phosphorylation

The addition or removal of phosphate groups from proteins is the most common chemical modification that regulates cellular processes. These modifications can affect the structure, activity, stability, and localization of proteins within cells as well as their interactions with other proteins.
During phosphorylation, protein kinases transfer the terminal phosphate group of ATP to specific amino acid side chains of substrate proteins. Serine, threonine, and tyrosine are the most commonly...

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by

Same author

Distinct IgM and IgG autoantibody profiles characterize incomplete and classified systemic autoimmune diseases.

Frontiers in immunology·2026

Same author

International expert consensus recommendations on standardised nomenclature of SSA/Ro (TROVE2/Ro60 and TRIM21/Ro52) autoantibodies in autoimmune diseases.

Annals of the rheumatic diseases·2026

Same author

Immune and stromal cell interaction in Sjögren's disease.

EBioMedicine·2026

Same author

A highly prevalent lupus risk haplotype increases IRF7-dependent induction of IFN-α, enhancing antiviral defense and exacerbating autoimmunity.

medRxiv : the preprint server for health sciences·2026

Same author

Muscarinic 3 receptor antibodies in Sjögren's disease: Evaluating assay variability and detection methods.

Journal of immunological methods·2026

Same author

There is no mechanism of sex bias in autoimmune disease: instead, there are mechanisms.

Rheumatology (Oxford, England)·2026

Search research articles

Related Experiment Video

Updated: Jun 23, 2026

Characterization at the Molecular Level using Robust Biochemical Approaches of a New Kinase Protein

Characterization at the Molecular Level using Robust Biochemical Approaches of a New Kinase Protein

Published on: June 30, 2019

Phosphoprotein detection on protein electroblot using a phosphate-specific fluorophore.

Lee Broderick Bockus¹, R Hal Scofield

¹Oklahoma Medical Research Foundation, 825 NE 13th Street, Oklahoma City, OK, 73104, USA. lee-bockus@omrf.org

Methods in Molecular Biology (Clifton, N.J.)

|April 21, 2009

Summary

A new fluorescent dye, Pro-Q Diamond, enables simple and reliable detection of phosphoproteins in blots. This technique offers selective phosphate group binding for accurate protein analysis.

More Related Videos

An Optimized Single-Molecule Pull-Down Assay for Quantification of Protein Phosphorylation

An Optimized Single-Molecule Pull-Down Assay for Quantification of Protein Phosphorylation

Published on: June 6, 2022

Related Experiment Videos

Last Updated: Jun 23, 2026

Characterization at the Molecular Level using Robust Biochemical Approaches of a New Kinase Protein

Characterization at the Molecular Level using Robust Biochemical Approaches of a New Kinase Protein

Published on: June 30, 2019

An Optimized Single-Molecule Pull-Down Assay for Quantification of Protein Phosphorylation

An Optimized Single-Molecule Pull-Down Assay for Quantification of Protein Phosphorylation

Published on: June 6, 2022

Area of Science:

Biochemistry
Molecular Biology
Proteomics

Background:

Phosphoprotein phosphorylation is a critical cellular regulatory mechanism.
Accurate detection of phosphoproteins is essential for understanding cellular pathways.

Purpose of the Study:

To develop a simple and reliable fluorescent detection technique for phosphoproteins in blots.
To adapt the Pro-Q Diamond dye for use in protein blotting applications.

Main Methods:

Utilized a small-molecule organic fluorophore, Pro-Q Diamond dye, for phosphoprotein detection.
Employed standard electrophoresis and electroblotting onto nitrocellulose or PVDF membranes.
Stained blots with Pro-Q Diamond dye, followed by destaining and imaging.

Main Results:

The Pro-Q Diamond dye selectively binds to the phosphate moiety of phosphoproteins.
Non-phosphorylated proteins, DNA, and RNA were not detected, ensuring specificity.
The technique is compatible with various imaging devices, MALDI-TOF-MS, and Edman sequencing.

Conclusions:

The Pro-Q Diamond dye offers a robust and versatile method for fluorescent detection of phosphoproteins in blots.
This technique simplifies phosphoprotein analysis and is compatible with downstream proteomic analyses.
The method enhances the study of phosphorylation-dependent cellular processes.