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

You might also read

Related Articles

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

Sort by

Same author

Optimizing cognitive enhancement through dopamine transporter modulation.

Neuropharmacology·2026

Same author

Sex and experience dependent regulation of synaptic protein turnover.

bioRxiv : the preprint server for biology·2025

Same author

Evaluation of PTSD-Induced Alterations in Bone Biomechanics and the Protective Potential of CE-123 in a Wistar Rat Model.

Journal of clinical medicine·2025

Same author

Reversal of the motivational effects of tetrabenazine by NMDA receptor blockade.

Neuropharmacology·2024

Same author

A brainstem-hypothalamus neuronal circuit reduces feeding upon heat exposure.

Nature·2024

Same author

Process Development and Scale-Up of a Novel Atypical DAT Inhibitor (<i>S</i>)-CE-123.

ACS omega·2024

Same journal

Author Correction: Novel octapeptide containing the RGD sequence as a potential anti-SARS-CoV-2 agent: design, synthesis, and theoretical studies.

Amino acids·2026

Same journal

Polyamines in CNS malignancies: positively charged culprits in the hijacking of neural and immune signaling pathways.

Amino acids·2026

Same journal

The effects of glutamine supplementation on inflammatory and oxidative stress indices in chronic diseases: a systematic review.

Amino acids·2026

Same journal

Reprogramming parasitic signatures into anticancer peptide candidates: in silico discovery of Leishmania major-derived peptides.

Amino acids·2026

Same journal

Effects of taurine on skeletal muscle in senescence-accelerated mice and C2C12 myoblasts and myotubes.

Amino acids·2026

Same journal

Dual functions of thermospermine in plant growth and stress responses.

Amino acids·2026

See all related articles

Search research articles

Related Experiment Video

Updated: Jan 18, 2026

Preparation of Naringenin Solution for In Vivo Application

Preparation of Naringenin Solution for In Vivo Application

Published on: August 10, 2021

In-gel total protein quantification using a ninhydrin-based method.

Sung Ung Kang¹, Seok Heo, Gert Lubec

¹Department of Pediatrics, Medical University of Vienna, Währinger Gürtel 18, 1090, Vienna, Austria, sungung.kang@gmail.com.

|July 10, 2013

Summary

This summary is machine-generated.

This study presents a reliable method for in-gel protein quantification using proteinase K digestion and ninhydrin reaction. The protocol accurately quantifies both hydrophilic and hydrophobic proteins, improving upon less reliable existing methods.

More Related Videos

Resin-Assisted Capture Coupled with Isobaric Tandem Mass Tag Labeling for Multiplexed Quantification of Protein Thiol Oxidation

Resin-Assisted Capture Coupled with Isobaric Tandem Mass Tag Labeling for Multiplexed Quantification of Protein Thiol Oxidation

Published on: June 21, 2021

Novel Whole-tissue Quantitative Assay of Nitric Oxide Levels in Drosophila Neuroinflammatory Response

Novel Whole-tissue Quantitative Assay of Nitric Oxide Levels in Drosophila Neuroinflammatory Response

Published on: December 4, 2013

Related Experiment Videos

Last Updated: Jan 18, 2026

Preparation of Naringenin Solution for In Vivo Application

Preparation of Naringenin Solution for In Vivo Application

Published on: August 10, 2021

Resin-Assisted Capture Coupled with Isobaric Tandem Mass Tag Labeling for Multiplexed Quantification of Protein Thiol Oxidation

Resin-Assisted Capture Coupled with Isobaric Tandem Mass Tag Labeling for Multiplexed Quantification of Protein Thiol Oxidation

Published on: June 21, 2021

Novel Whole-tissue Quantitative Assay of Nitric Oxide Levels in Drosophila Neuroinflammatory Response

Novel Whole-tissue Quantitative Assay of Nitric Oxide Levels in Drosophila Neuroinflammatory Response

Published on: December 4, 2013

Area of Science:

Biochemistry
Molecular Biology
Proteomics

Background:

Accurate in-gel protein quantification is crucial for downstream analyses.
Existing methods for relative protein quantification are often unreliable due to variations in protein migration and staining intensity.

Purpose of the Study:

To develop a precise and reliable method for quantifying both hydrophilic and hydrophobic proteins directly within gels.
To provide a protocol compatible with mass spectrometry-based protein identification.

Main Methods:

In-gel digestion of proteins using proteinase K.
Extraction and acid hydrolysis of digested proteins.
Quantification using the ninhydrin reaction.
Validation using bovine serum albumin as a standard on SDS-PAGE and IEF/SDS-PAGE gels.
Quantification of hydrophobic bacteriorhodopsin on SDS-PAGE gel.

Main Results:

The developed method provides accurate and reproducible in-gel protein quantification.
The protocol is suitable for both hydrophilic and hydrophobic proteins.
The method is compatible with subsequent mass spectrometric analysis.
Bovine serum albumin and bacteriorhodopsin were successfully quantified.

Conclusions:

The described protocol offers a reliable alternative for in-gel protein quantification beyond arbitrary optical density units.
This method enhances the accuracy of protein quantification for various biochemical and molecular analyses.
While time-consuming (4-7 days), the protocol ensures high accuracy and compatibility with mass spectrometry.