Immunoprecipitation of worm proteins

Worms expressing HA::RNP-6, RBM-39::mKate2 or both were harvested and proteins were extracted using the following standard protocol^{74 (link)}. A solubilization buffer containing 0.5% NP40, 150 mM NaCl and 50 mM Tris, pH 7.4 supplemented with cOmplete Protease Inhibitor (Roche) and PhosSTOP (Roche) was used for immunoprecipitation. Flag immunoprecipitation was performed using Dynabeads Protein G (Thermo Fisher Scientific) and FLAG M2 mouse monoclonal antibody (Sigma-Aldrich), following the manufacturers’ protocols. Proteins were eluted from the beads by boiling with Laemmli buffer.

Free full text: Click here

Huang W., Kew C., Fernandes S.D., Löhrke A., Han L., Demetriades C, & Antebi A. (2022). Decreased spliceosome fidelity and egl-8 intron retention inhibit mTORC1 signaling to promote longevity. Nature Aging, 2(9), 796-808.

Publication 2022

cOmplete Protease Inhibitor PhosSTOP Dynabeads Protein G FLAG M2 mouse monoclonal antibody

Buffer Immunoprecipitation Laemmli buffer Monoclonal antibody Mouse Nacl Protease inhibitor Protein g Proteins Tris Worms

Corresponding Organization :

Other organizations : Max Planck Institute for Biology of Ageing, Cologne Excellence Cluster on Cellular Stress Responses in Aging Associated Diseases, University of Cologne

Top 5 similar protocols

Variable analysis

independent variables

Worms expressing HA::RNP-6
Worms expressing RBM-39::mKate2
Worms expressing both HA::RNP-6 and RBM-39::mKate2

dependent variables

Proteins extracted from the worms

control variables

Solubilization buffer containing 0.5% NP40, 150 mM NaCl and 50 mM Tris, pH 7.4
COmplete Protease Inhibitor (Roche)
PhosSTOP (Roche)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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.

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?

Revolutionizing how scientists
search and build protocols!