The labeled peptides were fractionated by immobilized-pH-gradient isoelectric focusing (IPG-IEF), as previously described (17 (link),18 (link)). Briefly, the samples were dissolved in a Pharmalyte (GE Healthcare Life Sciences, Chalfont, UK) and urea solution, rehydrated on a pH 3–10 IPG strip, and subjected to IEF at 68 kV/h using an IPG phor system (GE Healthcare Life Sciences). The peptides were extracted from the gel using an acetonitrile (ACN) and formic acid solution (19 (link)). The fractions were lyophilized, and purified with SPE Discovery DSC-18 columns (Supelco, Inc., Bellefonte, PA, USA). The purified peptides were re-lyophilized and stored at −20°C until use.
Protocol detail
Peptide Fractionation by IPG-IEF
Partial Protocol Preview
This section provides a glimpse into the protocol.
The remaining content is hidden due to licensing restrictions, but the full text is available at the following link:
Access Free Full Text.
Corresponding Organization :
Other organizations : Dalian Medical University, Chongqing Medical University, Second Affiliated Hospital of Chongqing Medical University
Top 5 similar protocols
Variable analysis
independent variables
- Fractionation of labeled peptides by immobilized-pH-gradient isoelectric focusing (IPG-IEF)
- Peptide fractions extracted from the gel using an acetonitrile (ACN) and formic acid solution
- Purified peptides stored at -20°C
- Pharmalyte (GE Healthcare Life Sciences, Chalfont, UK) and urea solution used to dissolve samples
- PH 3–10 IPG strip used for rehydration and IEF
- IPG phor system (GE Healthcare Life Sciences) used for IEF at 68 kV/h
- SPE Discovery DSC-18 columns (Supelco, Inc., Bellefonte, PA, USA) used for peptide purification
Annotations
Based on most similar protocols
Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.
Sign up for free or login to display all annotations
As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!