In Vivo Chemical Cross-Linking with DSS

In vivo chemical cross-linking with disuccinimidyl suberate (DSS) (Thermo Fisher Scientific) was carried out as described [25 (link), 47 (link)] with some modifications. Cells transfected with expression plasmids for 17 h were washed three times in PBS with Ca²⁺/Mg²⁺ and then incubated with 1 mM DSS or DMSO (as control) in PBS for 1 h at room temperature. The reaction was stopped with a final concentration of 20 mM Tris (pH 7.5) for 15 min at room temperature. The samples were dissolved in NuPAGE^TM LDS sample buffer (4X) (Invitrogen) and analyzed by Western blotting.

Free full text: Click here

Yu R., Liu T., Jin S.B., Ankarcrona M., Lendahl U., Nistér M, & Zhao J. (2021). MIEF1/2 orchestrate mitochondrial dynamics through direct engagement with both the fission and fusion machineries. BMC Biology, 19, 229.

Publication 2021

disuccinimidyl suberate (DSS) NuPAGETM LDS sample buffer (4X)

Buffer Cells Disuccinimidyl suberate Dmso Plasmids Tris

Corresponding Organization :

Other organizations : Karolinska University Hospital, Karolinska Institutet

Top 5 similar protocols

Variable analysis

independent variables

Treatment with DSS (1 mM) or DMSO (control)

dependent variables

Western blot analysis of protein samples

control variables

Cells transfected with expression plasmids for 17 h
Cells washed three times in PBS with Ca2+/Mg2+
Reaction stopped with 20 mM Tris (pH 7.5) for 15 min at room temperature
Samples dissolved in NuPAGETM LDS sample buffer (4X)

negative control

DMSO treatment

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!