Overexpression and ELISA-based detection of asprosin in cultured cells and mice
Summary: 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 huma...
| Main Authors: | , |
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| Format: | Article |
| Language: | English |
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Elsevier
2022-12-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166722006426 |
| _version_ | 1811292317107093504 |
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| author | Ila Mishra Atul R. Chopra |
| author_facet | Ila Mishra Atul R. Chopra |
| author_sort | Ila Mishra |
| collection | DOAJ |
| description | Summary: 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). : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| first_indexed | 2024-04-13T04:43:37Z |
| format | Article |
| id | doaj.art-b011e27ff4894585a63d4e7202c48591 |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-04-13T04:43:37Z |
| publishDate | 2022-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-b011e27ff4894585a63d4e7202c485912022-12-22T03:01:55ZengElsevierSTAR Protocols2666-16672022-12-0134101762Overexpression and ELISA-based detection of asprosin in cultured cells and miceIla Mishra0Atul R. Chopra1Harrington Discovery Institute at University Hospitals, 2103 Cornell Road, Wolstein Research Building 4130, Cleveland, OH 44106, USAHarrington Discovery Institute at University Hospitals, 2103 Cornell Road, Wolstein Research Building 4130, Cleveland, OH 44106, USA; Department of Medicine, Case Western Reserve University, Cleveland, OH 44106, USA; Department of Genetics and Genome Sciences, Case Western Reserve University, Cleveland, OH 44106, USA; Corresponding authorSummary: 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). : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166722006426MetabolismMolecular biologyGene expressionAntibodyNeuroscienceMolecular/Chemical probes |
| spellingShingle | Ila Mishra Atul R. Chopra Overexpression and ELISA-based detection of asprosin in cultured cells and mice STAR Protocols Metabolism Molecular biology Gene expression Antibody Neuroscience Molecular/Chemical probes |
| title | Overexpression and ELISA-based detection of asprosin in cultured cells and mice |
| title_full | Overexpression and ELISA-based detection of asprosin in cultured cells and mice |
| title_fullStr | Overexpression and ELISA-based detection of asprosin in cultured cells and mice |
| title_full_unstemmed | Overexpression and ELISA-based detection of asprosin in cultured cells and mice |
| title_short | Overexpression and ELISA-based detection of asprosin in cultured cells and mice |
| title_sort | overexpression and elisa based detection of asprosin in cultured cells and mice |
| topic | Metabolism Molecular biology Gene expression Antibody Neuroscience Molecular/Chemical probes |
| url | http://www.sciencedirect.com/science/article/pii/S2666166722006426 |
| work_keys_str_mv | AT ilamishra overexpressionandelisabaseddetectionofasprosininculturedcellsandmice AT atulrchopra overexpressionandelisabaseddetectionofasprosininculturedcellsandmice |