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

Search research articles

Related Experiment Video

Updated: May 11, 2026

Lipid Droplet Isolation for Quantitative Mass Spectrometry Analysis

Lipid Droplet Isolation for Quantitative Mass Spectrometry Analysis

Published on: April 17, 2017

Lipid-binding analysis using a fat blot assay.

Teun Munnik¹, Magdalena Wierzchowiecka

¹Swammerdam Institute for Life Sciences, Section Plant Physiology, University of Amsterdam, Amsterdam, The Netherlands.

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

|May 18, 2013

Summary

This summary is machine-generated.

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

Acute remodeling of phosphoinositide lipids promotes endocytosis downstream of RALF/FERONIA signaling.

Current biology : CB·2026

Same author

Plasma membrane lipids at plant-pathogen interfaces: Regulators of immunity and susceptibility.

Progress in lipid research·2026

Same author

Molecular heaters: a green route to boosting crop yields?

Physical chemistry chemical physics : PCCP·2025

Same author

Plant PI-PLC signaling in stress and development.

Plant physiology·2025

Same author

Phosphate promotes Arabidopsis root skewing and circumnutation through reorganisation of the microtubule cytoskeleton.

The New phytologist·2024

Same author

Dual phosphorylation of DGK5-mediated PA burst regulates ROS in plant immunity.

Cell·2024

The Fat Blot assay is a simple, cost-effective method to study protein-lipid interactions. This technique uses immobilized lipids on membranes to measure protein binding affinity for specific lipids, aiding research in lipid metabolism and cell signaling.

Area of Science:

Biochemistry
Cell Biology
Molecular Biology

Background:

Protein-lipid interactions are crucial for cellular processes like metabolism, membrane trafficking, and signaling.
Understanding these interactions requires reliable and accessible experimental methods.

Purpose of the Study:

To describe and validate the Fat Blot assay for studying protein-lipid interactions.
To provide a simple, inexpensive method for qualitative and quantitative analysis of protein-lipid binding affinities.

Main Methods:

Lipids are immobilized via serial dilutions onto nitrocellulose membranes.
Lipid-binding proteins (e.g., GST-tagged) are incubated with the membranes.
Protein binding is detected using immunoblotting with specific antibodies.

More Related Videos

Lipid Vesicle-mediated Affinity Chromatography using Magnetic Activated Cell Sorting (LIMACS): a Novel Method to Analyze Protein-lipid Interaction

Lipid Vesicle-mediated Affinity Chromatography using Magnetic Activated Cell Sorting (LIMACS): a Novel Method to Analyze Protein-lipid Interaction

Published on: April 26, 2011

Shotgun Lipidomics of Rodent Tissues

Shotgun Lipidomics of Rodent Tissues

Published on: November 18, 2022

Related Experiment Videos

Last Updated: May 11, 2026

Lipid Droplet Isolation for Quantitative Mass Spectrometry Analysis

Lipid Droplet Isolation for Quantitative Mass Spectrometry Analysis

Published on: April 17, 2017

Lipid Vesicle-mediated Affinity Chromatography using Magnetic Activated Cell Sorting (LIMACS): a Novel Method to Analyze Protein-lipid Interaction

Lipid Vesicle-mediated Affinity Chromatography using Magnetic Activated Cell Sorting (LIMACS): a Novel Method to Analyze Protein-lipid Interaction

Published on: April 26, 2011

Shotgun Lipidomics of Rodent Tissues

Shotgun Lipidomics of Rodent Tissues

Published on: November 18, 2022

Main Results:

The Fat Blot assay allows for the assessment of relative binding affinities between proteins and lipids.
The method is adaptable for various lipid-binding proteins and lipid types.
The assay requires minimal protein input (micrograms) and is time-efficient.

Conclusions:

The Fat Blot assay is a valuable tool for investigating protein-lipid interactions.
Its simplicity, low cost, and efficiency make it suitable for diverse research applications.
This method facilitates the study of lipid-mediated cellular functions.