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Protocol to detect and quantify interactions between proteins expressed in Drosophila S2 cells.

Authors :
DeOliveira CC
Lin C
Crane BR
Source :
STAR protocols [STAR Protoc] 2024 Sep 20; Vol. 5 (3), pp. 103171. Date of Electronic Publication: 2024 Jul 05.
Publication Year :
2024

Abstract

Here, we present a protocol to quantify interactions among difficult-to-express proteins from Drosophila cells using the select western blot-free tagged-protein interaction (SWFTI) assay. We describe steps for plasmid design, cell plating, protein expression, and immunoprecipitation preparation. We then detail procedures for protein labeling, gel purification, and protein quantification. This protocol offers a fluorescence-based technique for rapid quantification of ectopically expressed proteins that are fused to SNAP and CLIP tags without the need for membrane transfer. For complete details on the use and execution of this protocol, please refer to Lin et al. <superscript>1</superscript> .<br />Competing Interests: Declaration of interests The authors declare no competing interests.<br /> (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Details

Language :
English
ISSN :
2666-1667
Volume :
5
Issue :
3
Database :
MEDLINE
Journal :
STAR protocols
Publication Type :
Academic Journal
Accession number :
38970794
Full Text :
https://doi.org/10.1016/j.xpro.2024.103171