Improve Research Reproducibility A Bio-protocol resource
Concise Method
tooltip

EphA2 gene knockdown using siRNA

HK Hee Sung Kim
YW You Jin Won
JS Ju Hee Shim
HK Hyun Ji Kim
JK Jihun Kim
HH Hea Nam Hong
BK Byung Sik Kim
request Request a Protocol
ask Ask a question
Favorite

siRNA for EphA2 was obtained from Santa Cruz (sc-29304; Santa Cruz Biotechnology, CA, USA). The target sequence was 5′-AATGACATGCCGATCTACATG-3′ (EphA2)43, and non-silencing siRNA sequence 5′-AATTCTCCGAACGTGTCACGT-3′ was used as negative control. AGS cells (5 × 105 cells/well) were transfected with siRNA at a final concentration of 20 to 100 nM using lipofectamine-2000 (Invitrogen). Following 8 h of transfection, cells were replaced with fresh medium containing 10% serum. Forty-eight hours after siRNA transfection, suppression of EphA2 expression was confirmed by western blot.

Copyright and License information: The Author(s) ©2019
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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, 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