Protocol for identifying components of subcellular compartments by antibody-based in situ biotinylation

Summary: Recent studies revealed that membrane-less subcellular organelles play important roles in cellular functions. Here, we present a protocol for identifying subcellular compartment components by antibody-based in situ biotinylation. We describe steps for in situ biotinylation labeling using a...

Full description

Saved in:
Bibliographic Details
Main Authors: Hidefumi Suzuki, Kexin Li, Sayaka Ito, Keiichiro Asano, Keisuke Noguchi, Ryota Abe, Hidehisa Takahashi
Format: Article
Language:English
Published: Elsevier 2025-06-01
Series:STAR Protocols
Subjects:
Online Access:http://www.sciencedirect.com/science/article/pii/S2666166725002096
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Summary: Recent studies revealed that membrane-less subcellular organelles play important roles in cellular functions. Here, we present a protocol for identifying subcellular compartment components by antibody-based in situ biotinylation. We describe steps for in situ biotinylation labeling using a horseradish peroxidase (HRP)-conjugated antibody, purification of the biotinylated components, and sample preparation for high-throughput analysis. This protocol has potential for application in the comprehensive analysis of dynamic subcellular organelles.For complete details on the use and execution of this protocol, please refer to Noguchi et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
ISSN:2666-1667