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Updated: Jun 17, 2026

Protein Complex Affinity Capture from Cryomilled Mammalian Cells

Protein Complex Affinity Capture from Cryomilled Mammalian Cells

Published on: December 9, 2016

Refined affinity purification protocol for CAMs using minimal mouse brain material.

Agathe Henocq¹, Wouter Doff², Dick Dekkers²

¹Department of Bionanoscience, Kavli Institute of Nanoscience Delft, Delft University of Technology, van der Maasweg 9, Delft 2629HZ, the Netherlands.

Journal of Neuroscience Methods

|February 25, 2026

Summary

This summary is machine-generated.

This study introduces a new protocol for identifying cell adhesion molecule (CAM) interactions using minimal mouse brain tissue. The refined method efficiently captures protein partners, aiding research into synaptic connectivity.

Keywords:

Animal refinement Neuronal surface Proteome Pull-down proteomics Synapse biology Synaptic cell adhesion

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Last Updated: Jun 17, 2026

Protein Complex Affinity Capture from Cryomilled Mammalian Cells

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Published on: December 9, 2016

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Subcellular Fractionation for the Isolation of Synaptic Components from the Murine Brain

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Published on: September 14, 2022

Area of Science:

Neuroscience
Molecular Biology
Biochemistry

Background:

Cell adhesion molecules (CAMs) are crucial for cell-cell interactions and play vital roles in neuronal development, wiring, and synaptic plasticity.
Identifying specific CAMs within dynamic macromolecular complexes is challenging due to transient interactions and difficulties in protein extraction.

Purpose of the Study:

To develop an accessible protocol for studying cell adhesion molecule interactions.
To enable the identification of CAM interacting partners from limited biological samples, specifically mouse brain lysate.

Main Methods:

A refined affinity purification protocol using a Teneurin-3-GFP bait protein was employed.
The method was optimized using minimal mouse brain lysate, comparing different detergents for lysate preparation.
Mass spectrometry was used to identify captured prey proteins, and normalization workflows were evaluated.

Main Results:

The protocol successfully detected known Teneurin interactants.
The refined approach significantly reduced the amount of animal tissue required compared to standard methods.
Different detergents impacted the enrichment of CAM species, highlighting the importance of lysate preparation.

Conclusions:

The developed protocol offers an accessible method for investigating CAM interactions.
This approach facilitates the study of complex protein networks underlying synaptic connectivity with reduced sample requirements.