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

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.

Capillary Electrophoresis: Applications

Capillary Electrophoresis: Applications

Capillary electrophoretic separations offer various modes, each with unique applications. These modes include capillary zone electrophoresis, capillary gel electrophoresis, capillary array electrophoresis, capillary isoelectric focusing, capillary isotachophoresis, micellar electrokinetic chromatography, and capillary electrochromatography.
Capillary zone electrophoresis (CZE) separates ionic components based on their electrophoretic mobility. It has been used to separate proteins, amino acids,...

You might also read

Related Articles

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

Sort by

Same author

Interrogating the dependency of AAV capsids on AAVR for retinal transduction.

Molecular therapy. Advances·2026

Same author

Laterally spreading AAV.SPR enables safe and efficient RS1 delivery to the macula after peripheral subretinal injection.

Molecular therapy : the journal of the American Society of Gene Therapy·2026

Same author

Correction: Inhibitor of Apoptosis-Stimulating Protein of p53 (iASPP) Is Required for Neuronal Survival after Axonal Injury.

PloS one·2026

Same author

Development of an AAV-based gene therapy for the ocular phenotype of Friedreich's ataxia.

Molecular therapy : the journal of the American Society of Gene Therapy·2025

Same author

A novel multiplex RNAi therapy simultaneously targets Hif1a and Hif2a to defy retinal degeneration in two models of AMD.

Molecular therapy : the journal of the American Society of Gene Therapy·2025

Same author

Assessment of the Orbit® subretinal delivery system (Orbit® SDS device) and prototypes in adult and juvenile canine eyes.

Drug delivery and translational research·2025

Same journal

Peptidomics in the Spotlight: Advanced Sample Treatment Techniques and Analytical Insights.

Advances in experimental medicine and biology·2026

Same journal

Methods for the Investigation of Protein-Ligands Interactions.

Advances in experimental medicine and biology·2026

Same journal

Sample Preparation Strategies for Microbial Cell Surface Proteomics: Integrating Shaving and Shotgun Approaches.

Advances in experimental medicine and biology·2026

Same journal

Proteomic Sample Preparation for the Petroleum Industry: A Biocorrosion Case Study.

Advances in experimental medicine and biology·2026

Same journal

Proteomic and Functional Comparison of Extracellular Vesicles from Wild-Type and Lyn-Deficient Stromal Cells.

Advances in experimental medicine and biology·2026

Same journal

Proteomic Analysis of Histone Sequence Variants and Post-translationally Modified Forms.

Advances in experimental medicine and biology·2026

See all related articles

Search research articles

Related Experiment Video

Updated: Jul 23, 2025

Procedure and Key Optimization Strategies for an Automated Capillary Electrophoretic-based Immunoassay Method

Procedure and Key Optimization Strategies for an Automated Capillary Electrophoretic-based Immunoassay Method

Published on: September 10, 2017

Optimization of Capillary-Based Western Blotting for MYO7A.

Kaitlyn R Calabro¹, Sanford L Boye¹, Shannon E Boye²

¹Department of Pediatrics, University of Florida, Gainesville, FL, USA.

Advances in Experimental Medicine and Biology

|July 13, 2023

Summary

This summary is machine-generated.

Researchers optimized a new immunoassay for Myosin VIIA (MYO7A) detection, crucial for developing dual adeno-associated virus (AAV) gene therapies to treat Usher syndrome type 1B (USH1B) vision loss.

Keywords:

Gene therapy MYO7A USH1B Usher syndrome

More Related Videos

Ultra-High-Speed Western Blot using Immunoreaction Enhancing Technology

Ultra-High-Speed Western Blot using Immunoreaction Enhancing Technology

Published on: September 26, 2020

A Guide to Modern Quantitative Fluorescent Western Blotting with Troubleshooting Strategies

A Guide to Modern Quantitative Fluorescent Western Blotting with Troubleshooting Strategies

Published on: November 20, 2014

Related Experiment Videos

Last Updated: Jul 23, 2025

Procedure and Key Optimization Strategies for an Automated Capillary Electrophoretic-based Immunoassay Method

Procedure and Key Optimization Strategies for an Automated Capillary Electrophoretic-based Immunoassay Method

Published on: September 10, 2017

Ultra-High-Speed Western Blot using Immunoreaction Enhancing Technology

Ultra-High-Speed Western Blot using Immunoreaction Enhancing Technology

Published on: September 26, 2020

A Guide to Modern Quantitative Fluorescent Western Blotting with Troubleshooting Strategies

A Guide to Modern Quantitative Fluorescent Western Blotting with Troubleshooting Strategies

Published on: November 20, 2014

Area of Science:

Genetics
Ophthalmology
Molecular Biology

Background:

Usher syndrome type 1B (USH1B) is a severe genetic disorder affecting hearing, balance, and vision.
The large size of the MYO7A gene poses challenges for gene therapy delivery.
Dual adeno-associated virus (AAV) vectors are being explored for USH1B gene therapy.

Purpose of the Study:

To develop and optimize a sensitive immunoassay for detecting Myosin VIIA (MYO7A) protein.
To support the development of dual AAV vector-based gene therapies for USH1B.
To improve the quantification of MYO7A expression in gene therapy models.

Main Methods:

Adaptation of traditional western blot protocols to the Jess™ Simple Western System.
Optimization of immunoassay conditions for MYO7A detection.
Testing and validation in mouse eye and HEK293 cell lysate samples.

Main Results:

An optimized Simple Western protocol was established for MYO7A detection.
The assay demonstrated improved sensitivity and quantifiability compared to traditional methods.
Successful MYO7A detection was achieved in relevant biological samples (mouse eyes, HEK293 cells).

Conclusions:

The optimized Simple Western assay is a valuable tool for evaluating dual AAV gene therapy efficacy for USH1B.
This method facilitates accurate assessment of MYO7A protein expression in gene therapy research.
Improved immunoassay sensitivity aids in the development of treatments for USH1B-related vision loss.