Improve Research Reproducibility A Bio-protocol resource
Concise Method
tooltip

Purification of photo-oxidation products

JK Jiwoong Kwon
JP Jong-Seok Park
MK Minsu Kang
SC Soobin Choi
JP Jumi Park
GK Gyeong Tae Kim
CL Changwook Lee
SC Sangwon Cha
HR Hyun-Woo Rhee
SS Sang-Hee Shim
request Request a Protocol
ask Ask a question
Favorite

The photo-oxidation products were extracted by using different solubility of the compounds between aqueous and organic solvents. We added equivalent volume of chloroform (528730, Sigma Aldrich) into bleached holoUnaG, and then strongly vortexed the mixed sample. The UnaG proteins were denatured and aggregated as a white pellet under this condition, where the organic layer was gradually turned into yellow color indicating that the UV-absorbing products were dissolved in the organic layer. We gently took the organic layer and evaporated the solvent at 60 °C with vacuum pump. The resulting yellow pellet was primarily dissolved in dimethyl sulfoxide (D8418, Sigma Aldrich), and further diluted for five times with deionized water before analysis.

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 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.

0/150

tip Tips for asking effective questions

+ Description

Write a detailed description. Include all information that will help others answer your question including experimental processes, conditions, and relevant images.

post Post a Question
0 Q&A