Overexpression and ELISA-based detection of asprosin in cultured cells and mice

Ila Mishra; Atul R. Chopra

doi:10.1016/j.xpro.2022.101762

Overexpression and ELISA-based detection of asprosin in cultured cells and mice

Ila Mishra, Atul R. Chopra

Research output: Contribution to journal › Article › peer-review

2 Scopus citations

Abstract

The unreliability of commercial recombinant asprosin preparations and variability between asprosin detection assays have proven to be a bottleneck in experimental interpretation. This protocol describes the use of viral vectors and expression plasmid for overexpression and secretion of human asprosin to achieve sustained elevation of asprosin protein in mice and HEK293T cells without using recombinant proteins. This protocol also includes a sandwich ELISA using anti-asprosin monoclonal antibodies for detection of asprosin in media from cultured cells and in plasma of mice. For complete details on the use and execution of this protocol, please refer to Duerrschmid et al. (2017), Mishra et al. (2021), and Mishra et al. (2022).

Original language	English
Article number	101762
Journal	STAR Protocols
Volume	3
Issue number	4
DOIs	https://doi.org/10.1016/j.xpro.2022.101762
State	Published - Dec 16 2022

Bibliographical note

Publisher Copyright:
© 2022 The Author(s)

Funding

This work was supported by the NIDDK ( DK102529 , DK118290 ). We thank members of the Chopra lab for helpful suggestions and critical reading of the manuscript. We thank Zhiqiang An for rabbit monoclonal anti-asprosin antibody development. Summary figure created using Biorender.com . This work was supported by the NIDDK (DK102529, DK118290). We thank members of the Chopra lab for helpful suggestions and critical reading of the manuscript. We thank Zhiqiang An for rabbit monoclonal anti-asprosin antibody development. Summary figure created using Biorender.com. Investigation, methodology, and writing, I.M.; conceptualization, resources, supervision, funding acquisition, and writing – review and editing, A.R.C. A.R.C. has been awarded asprosin-related patents and is a co-founder, director, and officer of Vizigen, Inc. and Aceragen, Inc. and holds equity in both companies.

Funders	Funder number
A.R.C. A.R.C.
National Institute of Diabetes and Digestive and Kidney Diseases	DK118290, DK102529

Keywords

Antibody
Gene expression
Metabolism
Molecular biology
Molecular/Chemical probes
Neuroscience

ASJC Scopus subject areas

General Neuroscience
General Biochemistry, Genetics and Molecular Biology
General Immunology and Microbiology

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10.1016/j.xpro.2022.101762

Cite this

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abstract = "The unreliability of commercial recombinant asprosin preparations and variability between asprosin detection assays have proven to be a bottleneck in experimental interpretation. This protocol describes the use of viral vectors and expression plasmid for overexpression and secretion of human asprosin to achieve sustained elevation of asprosin protein in mice and HEK293T cells without using recombinant proteins. This protocol also includes a sandwich ELISA using anti-asprosin monoclonal antibodies for detection of asprosin in media from cultured cells and in plasma of mice. For complete details on the use and execution of this protocol, please refer to Duerrschmid et al. (2017), Mishra et al. (2021), and Mishra et al. (2022).",

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Overexpression and ELISA-based detection of asprosin in cultured cells and mice

Abstract

Bibliographical note

Funding

Keywords

ASJC Scopus subject areas

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