Improve Research Reproducibility A Bio-protocol resource
Concise Method
tooltip

Co-immunoprecipitation

KS Karolina Serwach
EN Ewa Nurowska
MK Marta Klukowska
BZ Barbara Zablocka
JG Joanna Gruszczynska-Biegala
request Request a Protocol
ask Ask a question
Favorite

For the Co-IP of proteins, neurons were collected and lysed for 1.5 h. Lysates were centrifuged at 15,000 × g for 15 min at 4 °C, and the supernatant was collected and mixed with Protein G-Agarose beads (Roche). Precleared lysates (700 µg) were incubated overnight with protein G-Agarose that was preincubated for 3 h with the following antibodies: rabbit anti-GluN2A (Alomone, Catalog No. AGC-002) or rabbit anti-GluN2B (Proteintech, Catalog No. 21920-1-AP). Rabbit anti-immunoglobulin G (IgG) antibody (Merck, Catalog No. 12-370 and 12-371) was used as a negative control. The precipitated proteins were eluted and separated by SDS-PAGE.

Copyright and License information: The Author(s) ©2023
Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

Do you have any questions about this protocol?

Post your question to gather feedback from the community. We will also invite the authors of this article to respond.

post Post a Question
0 Q&A