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-Drug Binding: Determination Methods

Protein-Drug Binding: Determination Methods

Determining protein-drug binding can be achieved through indirect and direct methods, each providing valuable insights into the interaction between proteins and drugs.
Indirect methods involve isolating the bound drug from its free form in biological samples such as blood, serum, or plasma. These techniques aim to measure the percentage of drugs bound to proteins. Equilibrium dialysis is a commonly used method where the free drug concentration at equilibrium is measured by separating the bound...

You might also read

Related Articles

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

Sort by

Same author

A dual sensor regulates P-glycoprotein's structural plasticity.

bioRxiv : the preprint server for biology·2026

Same author

SF3B1 Phosphorylation Prompts U2AF2 Dissociation for Widespread Control of pre-mRNA Splicing.

bioRxiv : the preprint server for biology·2026

Same author

Molecular Phenotypes Segregate Missense Mutations in SLC13A5 Epilepsy.

Journal of molecular biology·2024

Same author

Recurrent Neurodevelopmentally Associated Variants of the Pre-mRNA Splicing Factor U2AF2 Alter RNA Binding Affinities and Interactions.

Biochemistry·2024

Same author

Molecular phenotypes segregate missense mutations in SLC13A5 Epilepsy.

bioRxiv : the preprint server for biology·2024

Same author

Lipid environment determines the drug-stimulated ATPase activity of P-glycoprotein.

Frontiers in molecular biosciences·2023

Search research articles

Related Experiment Video

Updated: Dec 25, 2025

Microvolume Protein Concentration Determination using the NanoDrop 2000c Spectrophotometer

Microvolume Protein Concentration Determination using the NanoDrop 2000c Spectrophotometer

Published on: November 4, 2009

Methods for Measuring the Concentrations of Proteins.

Clara L Kielkopf, William Bauer, Ina L Urbatsch

Cold Spring Harbor Protocols

|April 3, 2020

Summary

Protein concentration is typically measured using UV absorbance or colorimetric assays like Bradford. Both methods have limitations, so comparing results from multiple assays is recommended for critical applications.

Area of Science:

Biochemistry
Analytical Chemistry
Protein Analysis

Background:

Accurate protein concentration determination is crucial for biochemical and molecular biology experiments.
Common methods include UV absorbance at 280 nm (A280) and colorimetric assays (Bradford, Lowry, BCA).

Purpose of the Study:

To evaluate the advantages and limitations of different protein concentration determination methods.
To provide guidance on selecting appropriate methods based on experimental needs and potential interferences.

Main Methods:

Review of established protein quantification techniques, focusing on UV absorbance (A280) and colorimetric assays.
Discussion of inherent constraints and potential interferences associated with each method.

More Related Videos

Selected Reaction Monitoring Mass Spectrometry for Absolute Protein Quantification

Selected Reaction Monitoring Mass Spectrometry for Absolute Protein Quantification

Published on: August 17, 2015

Robust Comparison of Protein Levels Across Tissues and Throughout Development Using Standardized Quantitative Western Blotting

Robust Comparison of Protein Levels Across Tissues and Throughout Development Using Standardized Quantitative Western Blotting

Published on: April 9, 2019

Related Experiment Videos

Last Updated: Dec 25, 2025

Microvolume Protein Concentration Determination using the NanoDrop 2000c Spectrophotometer

Microvolume Protein Concentration Determination using the NanoDrop 2000c Spectrophotometer

Published on: November 4, 2009

Selected Reaction Monitoring Mass Spectrometry for Absolute Protein Quantification

Selected Reaction Monitoring Mass Spectrometry for Absolute Protein Quantification

Published on: August 17, 2015

Robust Comparison of Protein Levels Across Tissues and Throughout Development Using Standardized Quantitative Western Blotting

Robust Comparison of Protein Levels Across Tissues and Throughout Development Using Standardized Quantitative Western Blotting

Published on: April 9, 2019

Main Results:

UV absorbance (A280) is fast, convenient, and non-consumptive for purified proteins but susceptible to buffer and contaminant interference.
Colorimetric assays (Bradford, Lowry, BCA) offer alternatives but can be affected by protein composition and buffer reagents.
No single method is universally perfect; each has specific limitations.

Conclusions:

For purified proteins, A280 is often preferred due to its simplicity and non-destructive nature.
Colorimetric assays are valuable but require careful consideration of reagent compatibility and protein variability.
For critical applications, such as enzyme kinetics, cross-validation using multiple protein quantification methods is advisable.