MAD1 peptide was synthesized as described previously.[9 (link)] In brief, Fmoc-based solid-phase peptide synthesis was performed on Rink Amide ProTide Resin using CEM Liberty Blue Automated Microwave Peptide Synthesizer. Fluorescent labeling of MAD1 using NHS-Fluorescein (FITC-MAD1) was performed as previously described.[31 (link)] Next, peptide cleavage and deprotection was performed by stirring in TFA:thioanisole:1,2-ethanedithiol:anisole (90:5:3:2 ratio) solution under argon for 3 hours at 40°C. Peptides were purified by reverse-phase HPLC (Shimadzu; Columbia, MD) using a Phenomenex Luna Omega PS C18 column (Torrance, CA) with a linear 1%/min. solvent gradient of solvent A (0.1% TFA) and solvent B (0.1% TFA in 90% acetonitrile). Purity of MAD1 peptide was confirmed by reverse-phase HPLC-MS (Figure S9 ). Purified MAD1 was lyophilized and stored at −20 °C before use.
Protocol detail
Synthesis and Purification of MAD1 Peptide
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.
Acetonitrile
Anisole
Argon
Cleavage
Ethanedithiol
Fitc
Fluorescein
Hplc
Microwave
Peptide
Rink amide resin
Solvent
Thioanisole
Corresponding Organization :
Other organizations : Pennsylvania State University, Center for Disease Dynamics, Economics & Policy
Top 5 similar protocols
Variable analysis
independent variables
- Fmoc-based solid-phase peptide synthesis
- Fluorescent labeling of MAD1 using NHS-Fluorescein (FITC-MAD1)
- Peptide cleavage and deprotection in TFA:thioanisole:1,2-ethanedithiol:anisole (90:5:3:2 ratio) solution under argon for 3 hours at 40°C
- Reverse-phase HPLC purification of peptides
- Purity of MAD1 peptide
- Rink Amide ProTide Resin used for solid-phase peptide synthesis
- CEM Liberty Blue Automated Microwave Peptide Synthesizer used for solid-phase peptide synthesis
- Phenomenex Luna Omega PS C18 column used for reverse-phase HPLC purification
- Lyophilization and storage at -20°C before use
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!