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Journal Article

MISTIC-fusion proteins as antigens for high quality membrane protein antibodies

MPS-Authors
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Silva Alves,  N
Antonin Group, Friedrich Miescher Laboratory, Max Planck Society;

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Astrinidis,  SA
Antonin Group, Friedrich Miescher Laboratory, Max Planck Society;

/persons/resource/persons272488

Eisenhardt,  N
Antonin Group, Friedrich Miescher Laboratory, Max Planck Society;

/persons/resource/persons32702

Sieverding,  C
Antonin Group, Friedrich Miescher Laboratory, Max Planck Society;

/persons/resource/persons272491

Redolfi,  J
Antonin Group, Friedrich Miescher Laboratory, Max Planck Society;

/persons/resource/persons94125

Lorenz,  M
Antonin Group, Friedrich Miescher Laboratory, Max Planck Society;

/persons/resource/persons271957

Weberruss,  M
Antonin Group, Friedrich Miescher Laboratory, Max Planck Society;

/persons/resource/persons271960

Moreno-Andres,  D
Antonin Group, Friedrich Miescher Laboratory, Max Planck Society;

/persons/resource/persons14783

Antonin,  W
Antonin Group, Friedrich Miescher Laboratory, Max Planck Society;

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Citation

Silva Alves, N., Astrinidis, S., Eisenhardt, N., Sieverding, C., Redolfi, J., Lorenz, M., et al. (2017). MISTIC-fusion proteins as antigens for high quality membrane protein antibodies. Scientific Reports, 7: 41519. doi:10.1038/srep41519.


Cite as: https://hdl.handle.net/21.11116/0000-000A-5527-B
Abstract
Lack of high-quality antibodies against transmembrane proteins is a widely recognized hindrance in biomedical and cell biological research. Here we present a robust pipeline for the generation of polyclonal antibodies employing full-length membrane proteins as immunogens to overcome this "antibody bottleneck". We express transmembrane proteins fused to a MISTIC fragment that enhances expression of eukaryotic membrane proteins in E. coli. Purified membrane proteins are used as immunogen for rabbit injection employing standard immunizing protocols. The raised antibodies against membrane proteins of the endoplasmic reticulum and the nuclear envelope, which we use as test cases, function in a wide range of applications and are superior to ones produced against soluble domains as immunogens.