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

Calreticulin-targeting trisaccharide Glc-Man-Man conjugated with zinc chlorophyll derivative as a potential photosensitizer.

Photochemistry and photobiology·2026

Same author

INHAND: Non-proliferative and proliferative lesions of fish - Chapter 11. Fish respiratory system.

Journal of toxicologic pathology·2026

Same author

INHAND: Non-proliferative and proliferative lesions of fish - Chapter 4. Fish endocrine system.

Journal of toxicologic pathology·2026

Same author

INHAND: Non-proliferative and proliferative lesions of fish - Chapter 8. Fish nervous system.

Journal of toxicologic pathology·2026

Same author

INHAND: Non-proliferative and proliferative lesions of fish - Chapter 14. Fish special senses.

Journal of toxicologic pathology·2026

Same author

INHAND: Non-proliferative and proliferative lesions of fish - Chapter 7. Fish integumentary system.

Journal of toxicologic pathology·2026

Search research articles

Related Experiment Video

Updated: Apr 20, 2026

In Vitro Characterization of Histone Chaperones using Analytical, Pull-Down and Chaperoning Assays

In Vitro Characterization of Histone Chaperones using Analytical, Pull-Down and Chaperoning Assays

Published on: December 29, 2021

Analytical method for determining relative chaperone activity using an ovalbumin-conjugated column.

Makoto Hirano¹, Yuki Kato¹, Ayami Imagawa¹

¹Department of Materials and Life Science, Seikei University, 3-3-1 Kichijoji-kitamachi, Musashino-shi, Tokyo 180-8633, Japan.

Biochemical and Biophysical Research Communications

|December 2, 2014

Summary

This study introduces a new method to measure molecular chaperone activity using a protein-conjugated column. The assay revealed that calreticulin exhibits stronger binding to denatured ovalbumin than BiP.

Keywords:

BiP Calreticulin Molecular chaperone OVA-conjugated column Relative activity

More Related Videos

Detection of the pH-dependent Activity of Escherichia coli Chaperone HdeB In Vitro and In Vivo

Detection of the pH-dependent Activity of Escherichia coli Chaperone HdeB In Vitro and In Vivo

Published on: October 23, 2016

Studies of Chaperone-Cochaperone Interactions using Homogenous Bead-Based Assay

Studies of Chaperone-Cochaperone Interactions using Homogenous Bead-Based Assay

Published on: July 21, 2021

Related Experiment Videos

Last Updated: Apr 20, 2026

In Vitro Characterization of Histone Chaperones using Analytical, Pull-Down and Chaperoning Assays

In Vitro Characterization of Histone Chaperones using Analytical, Pull-Down and Chaperoning Assays

Published on: December 29, 2021

Detection of the pH-dependent Activity of Escherichia coli Chaperone HdeB In Vitro and In Vivo

Detection of the pH-dependent Activity of Escherichia coli Chaperone HdeB In Vitro and In Vivo

Published on: October 23, 2016

Studies of Chaperone-Cochaperone Interactions using Homogenous Bead-Based Assay

Studies of Chaperone-Cochaperone Interactions using Homogenous Bead-Based Assay

Published on: July 21, 2021

Area of Science:

Biochemistry
Molecular Biology
Cellular Biology

Background:

Understanding protein biosynthesis and the role of molecular chaperones is crucial for cell function.
Existing methods for assessing chaperone activity may not accurately reflect physiological conditions with multiple chaperones present.

Purpose of the Study:

To develop and validate an analytical method for estimating relative molecular chaperone activity under physiological, multi-chaperone conditions.
To compare the relative affinities of different molecular chaperones for a denatured protein using a novel chromatography-based assay.

Main Methods:

Development of a protein-conjugated column assay using denatured ovalbumin.
Subjecting chaperone mixtures to chromatography on the conjugated column.
Utilizing western blotting to determine elution positions and infer relative chaperone affinity and activity.

Main Results:

The developed analytical method effectively estimated relative chaperone activity.
In a mixture of calreticulin and BiP, calreticulin demonstrated a stronger association with denatured ovalbumin compared to BiP.
Elution position correlated with chaperone activity, validating the method's principle.

Conclusions:

The protein-conjugated column assay is a viable method for assessing relative molecular chaperone activity in complex mixtures.
Calreticulin possesses higher affinity for denatured ovalbumin than BiP under the tested conditions.
This method advances the understanding of chaperone interactions and protein homeostasis within the cell.