A guide to selecting high-performing antibodies for Serine/threonine-protein phosphatase 2A 56 kDa regulatory subunit delta isoform (PPP2R5D) for use in Western Blot, immunoprecipitation and immunofluorescence [version 2; peer review: 2 approved]
Protein phosphatase 2A is a serine/threonine phosphatase with activity dependent on an associated regulatory subunit, serine/threonine-protein phosphatase 2A 56 kDa regulatory subunit delta (δ) isoform (PPP2R5D). PPP2R5D is the δ isoform in the B56 family of regulatory subunits. Abundantly expressed...
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F1000 Research Ltd
2024-07-01
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| author | Carl Laflamme Kathleen Southern Riham Ayoubi Maryam Fotouhi Charles Alende |
| author_facet | Carl Laflamme Kathleen Southern Riham Ayoubi Maryam Fotouhi Charles Alende |
| author_sort | Carl Laflamme |
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| description | Protein phosphatase 2A is a serine/threonine phosphatase with activity dependent on an associated regulatory subunit, serine/threonine-protein phosphatase 2A 56 kDa regulatory subunit delta (δ) isoform (PPP2R5D). PPP2R5D is the δ isoform in the B56 family of regulatory subunits. Abundantly expressed in the brain and involved in a broad range of cellular processes, PPP2R5D plays an essential role in modulating key neuronal pathways and signalling. Pathogenic mutations in the PPP2R5D gene are linked to clinical symptoms characterized by neurodevelopmental delay, intellectual disability, and autism spectrum disorders. The etiology of these genetic disorders remains unknown, which can partly be due to the lack of independently characterized antibodies. Here we have characterized six PPP2R5D commercial antibodies for Western Blot, immunoprecipitation, and immunofluorescence using a standardized experimental protocol based on comparing read-outs in knockout cell lines and isogenic parental controls. These studies are part of a larger, collaborative initiative seeking to address antibody reproducibility by characterizing commercially available antibodies for human proteins and publishing the results openly as a resource for the scientific community. While use of antibodies and protocols vary between laboratories, we encourage readers to use this report as a guide to select the most appropriate antibodies for their specific needs. |
| format | Article |
| id | doaj-art-1ee7f6a76f7a4e23b5574fe58cfa8eb6 |
| institution | Kabale University |
| issn | 2046-1402 |
| language | English |
| publishDate | 2024-07-01 |
| publisher | F1000 Research Ltd |
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| spelling | doaj-art-1ee7f6a76f7a4e23b5574fe58cfa8eb62025-02-11T01:00:01ZengF1000 Research LtdF1000Research2046-14022024-07-0113168890A guide to selecting high-performing antibodies for Serine/threonine-protein phosphatase 2A 56 kDa regulatory subunit delta isoform (PPP2R5D) for use in Western Blot, immunoprecipitation and immunofluorescence [version 2; peer review: 2 approved]Carl Laflamme0https://orcid.org/0000-0001-5906-025XKathleen Southern1https://orcid.org/0000-0002-4125-3608Riham Ayoubi2Maryam Fotouhi3Charles Alende4https://orcid.org/0009-0005-4611-6134Department of Neurology and Neurosurgery, Structural Genomics Consortium, The Montreal Neurological Institute, McGill University, Montreal, Québec, H3A 2B4, CanadaDepartment of Neurology and Neurosurgery, Structural Genomics Consortium, The Montreal Neurological Institute, McGill University, Montreal, Québec, H3A 2B4, CanadaDepartment of Neurology and Neurosurgery, Structural Genomics Consortium, The Montreal Neurological Institute, McGill University, Montreal, Québec, H3A 2B4, CanadaDepartment of Neurology and Neurosurgery, Structural Genomics Consortium, The Montreal Neurological Institute, McGill University, Montreal, Québec, H3A 2B4, CanadaDepartment of Neurology and Neurosurgery, Structural Genomics Consortium, The Montreal Neurological Institute, McGill University, Montreal, Québec, H3A 2B4, CanadaProtein phosphatase 2A is a serine/threonine phosphatase with activity dependent on an associated regulatory subunit, serine/threonine-protein phosphatase 2A 56 kDa regulatory subunit delta (δ) isoform (PPP2R5D). PPP2R5D is the δ isoform in the B56 family of regulatory subunits. Abundantly expressed in the brain and involved in a broad range of cellular processes, PPP2R5D plays an essential role in modulating key neuronal pathways and signalling. Pathogenic mutations in the PPP2R5D gene are linked to clinical symptoms characterized by neurodevelopmental delay, intellectual disability, and autism spectrum disorders. The etiology of these genetic disorders remains unknown, which can partly be due to the lack of independently characterized antibodies. Here we have characterized six PPP2R5D commercial antibodies for Western Blot, immunoprecipitation, and immunofluorescence using a standardized experimental protocol based on comparing read-outs in knockout cell lines and isogenic parental controls. These studies are part of a larger, collaborative initiative seeking to address antibody reproducibility by characterizing commercially available antibodies for human proteins and publishing the results openly as a resource for the scientific community. While use of antibodies and protocols vary between laboratories, we encourage readers to use this report as a guide to select the most appropriate antibodies for their specific needs.https://f1000research.com/articles/13-1/v2Uniprot ID Q14738 PPP2R5D Serine/threonine-protein phosphatase 2A 56 kDa regulatory subunit delta isoform antibody characterization antibody validation Western Bloteng |
| spellingShingle | Carl Laflamme Kathleen Southern Riham Ayoubi Maryam Fotouhi Charles Alende A guide to selecting high-performing antibodies for Serine/threonine-protein phosphatase 2A 56 kDa regulatory subunit delta isoform (PPP2R5D) for use in Western Blot, immunoprecipitation and immunofluorescence [version 2; peer review: 2 approved] F1000Research Uniprot ID Q14738 PPP2R5D Serine/threonine-protein phosphatase 2A 56 kDa regulatory subunit delta isoform antibody characterization antibody validation Western Blot eng |
| title | A guide to selecting high-performing antibodies for Serine/threonine-protein phosphatase 2A 56 kDa regulatory subunit delta isoform (PPP2R5D) for use in Western Blot, immunoprecipitation and immunofluorescence [version 2; peer review: 2 approved] |
| title_full | A guide to selecting high-performing antibodies for Serine/threonine-protein phosphatase 2A 56 kDa regulatory subunit delta isoform (PPP2R5D) for use in Western Blot, immunoprecipitation and immunofluorescence [version 2; peer review: 2 approved] |
| title_fullStr | A guide to selecting high-performing antibodies for Serine/threonine-protein phosphatase 2A 56 kDa regulatory subunit delta isoform (PPP2R5D) for use in Western Blot, immunoprecipitation and immunofluorescence [version 2; peer review: 2 approved] |
| title_full_unstemmed | A guide to selecting high-performing antibodies for Serine/threonine-protein phosphatase 2A 56 kDa regulatory subunit delta isoform (PPP2R5D) for use in Western Blot, immunoprecipitation and immunofluorescence [version 2; peer review: 2 approved] |
| title_short | A guide to selecting high-performing antibodies for Serine/threonine-protein phosphatase 2A 56 kDa regulatory subunit delta isoform (PPP2R5D) for use in Western Blot, immunoprecipitation and immunofluorescence [version 2; peer review: 2 approved] |
| title_sort | guide to selecting high performing antibodies for serine threonine protein phosphatase 2a 56 kda regulatory subunit delta isoform ppp2r5d for use in western blot immunoprecipitation and immunofluorescence version 2 peer review 2 approved |
| topic | Uniprot ID Q14738 PPP2R5D Serine/threonine-protein phosphatase 2A 56 kDa regulatory subunit delta isoform antibody characterization antibody validation Western Blot eng |
| url | https://f1000research.com/articles/13-1/v2 |
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