Improve Research Reproducibility A Bio-protocol resource
Concise Method
tooltip

Antigen presentation assay

YT Yuki Tanaka
HW Hiroshi Wada
RG Risa Goto
TO Toshihiro Osada
KY Keisuke Yamamura
SF Satoshi Fukaya
AS Atsushi Shimizu
MO Mitsuru Okubo
KM Kazuhisa Minamiguchi
KI Koichi Ikizawa
ES Eiji Sasaki
TU Teruhiro Utsugi
request Request a Protocol
ask Ask a question
Favorite

CD11c+ dendritic cells (DCs) and CD3+ T cells were purified with CD11c MicroBeads UltraPure, mouse, and the Pan T Cell Isolation Kit II, mouse (Miltenyi Biotec) from the splenocytes of HLA-A*0201 KI mice. The antigen-presenting cells were incubated with peptides for 3 h (37 °C, 5% CO2) and washed three times. Epoxomicin was added 30 min before the addition of TAS0314. Cultured SART3302–310-specific CTLs from immunized HLA-A*0201 KI mice with SART3302–310 (100 μg/mice) were used as effector cells. The CTLs were co-cultivated with antigen-presenting cells on an anti-mouse IFN-γ-coated plate. Epitope-specific IFN-γ production was detected using an ELISPOT assay.

Copyright and License information: The Author(s) ©2020
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