Improve Research Reproducibility A Bio-protocol resource
Concise Method
tooltip

WES

HL Hua Liu
JL Jianjiao Li
YY Ying Yang
LL Liu Liu
LY Lifu Yu
MT Minsong Tu
RY Ruihong Yuan
WY Wanyuan Yue
QL Qi Luo
YR Yonghua Ruan
XD Xiaoming Dai
request Request a Protocol
ask Ask a question
Favorite

Western blotting was performed using WES (ProteinSimple, San Jose, CA, USA). Briefly, 8 μL diluted protein lysate was mixed with 2 μL of 5× fluorescent master mix and heated at 95 °C for 5 min. The samples, blocking reagent, wash buffer, primary antibodies, secondary antibodies, and chemiluminescent substrate were dispensed into designated wells in a microplate. The plate was loaded into the instrument, and protein was drawn into individual capillaries on a 25-capillary cassette. Protein separation and chemiluminescence were performed automatically on individual capillaries. Data were analyzed using Compass software (ProteinSimple). Anti-CYLD rabbit mAb (8462 T, CST, Danvers, MA, USA) and beta-actin rabbit mAb (4970 S, CST) were used as primary antibody and loading control.

Copyright and License information: The Author(s) ©2018
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