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

Ila Mishra, Atul R. Chopra

Research output: Contribution to journalArticlepeer-review

1 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 languageEnglish
Article number101762
JournalSTAR Protocols
Volume3
Issue number4
DOIs
StatePublished - Dec 16 2022

Bibliographical note

Publisher Copyright:
© 2022 The Author(s)

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

Fingerprint

Dive into the research topics of 'Overexpression and ELISA-based detection of asprosin in cultured cells and mice'. Together they form a unique fingerprint.

Cite this