A guide to selecting high-performing antibodies for Huntingtin (UniProt ID: P42858) for use in western blot, immunoprecipitation, and immunofluorescence [version 2; peer review: 2 approved]
Huntingtin encodes a 3144 amino acid protein, with a polyglutamine repeat tract at the N-terminus. Expansion of this repeat tract above a pathogenic threshold of 36 repeats is the causative mutation of Huntington's disease, a neurodegenerative disorder characterized by loss of striatal neurons....
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2025-01-01
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| author | Rachel J. Harding Carl Laflamme Sara González Bolívar Aled M. Edwards Kathleen Southern Rebeka Fanti Riham Ayoubi Charles Alende Maryam Fotouhi Renu Chandrasekaran |
| author_facet | Rachel J. Harding Carl Laflamme Sara González Bolívar Aled M. Edwards Kathleen Southern Rebeka Fanti Riham Ayoubi Charles Alende Maryam Fotouhi Renu Chandrasekaran |
| author_sort | Rachel J. Harding |
| collection | DOAJ |
| description | Huntingtin encodes a 3144 amino acid protein, with a polyglutamine repeat tract at the N-terminus. Expansion of this repeat tract above a pathogenic threshold of 36 repeats is the causative mutation of Huntington's disease, a neurodegenerative disorder characterized by loss of striatal neurons. Here we have characterized twenty Huntingtin 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 issues 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-8354cdb4f0a14d4992484fa349d78c49 |
| institution | Kabale University |
| issn | 2046-1402 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | F1000 Research Ltd |
| record_format | Article |
| series | F1000Research |
| spelling | doaj-art-8354cdb4f0a14d4992484fa349d78c492025-01-03T01:00:04ZengF1000 Research LtdF1000Research2046-14022025-01-0113176211A guide to selecting high-performing antibodies for Huntingtin (UniProt ID: P42858) for use in western blot, immunoprecipitation, and immunofluorescence [version 2; peer review: 2 approved]Rachel J. Harding0https://orcid.org/0000-0002-1134-391XCarl Laflamme1https://orcid.org/0000-0001-5906-025XSara González Bolívar2https://orcid.org/0000-0002-4299-8281Aled M. Edwards3Kathleen Southern4https://orcid.org/0000-0002-4125-3608Rebeka Fanti5Riham Ayoubi6Charles Alende7https://orcid.org/0009-0005-4611-6134Maryam Fotouhi8Renu Chandrasekaran9Structural Genomics Consortium, University of Toronto, Toronto, Ontario, CanadaDepartment of Neurology and Neurosurgery, Structural Genomics Consortium, The Montreal Neurological Institute, McGill University, Montreal, Québec, CanadaDepartment of Neurology and Neurosurgery, Structural Genomics Consortium, The Montreal Neurological Institute, McGill University, Montreal, Québec, CanadaStructural Genomics Consortium, University of Toronto, Toronto, Ontario, CanadaDepartment of Neurology and Neurosurgery, Structural Genomics Consortium, The Montreal Neurological Institute, McGill University, Montreal, Québec, CanadaStructural Genomics Consortium, University of Toronto, Toronto, Ontario, CanadaDepartment of Neurology and Neurosurgery, Structural Genomics Consortium, The Montreal Neurological Institute, McGill University, Montreal, Québec, CanadaDepartment of Neurology and Neurosurgery, Structural Genomics Consortium, The Montreal Neurological Institute, McGill University, Montreal, Québec, CanadaDepartment of Neurology and Neurosurgery, Structural Genomics Consortium, The Montreal Neurological Institute, McGill University, Montreal, Québec, CanadaStructural Genomics Consortium, University of Toronto, Toronto, Ontario, CanadaHuntingtin encodes a 3144 amino acid protein, with a polyglutamine repeat tract at the N-terminus. Expansion of this repeat tract above a pathogenic threshold of 36 repeats is the causative mutation of Huntington's disease, a neurodegenerative disorder characterized by loss of striatal neurons. Here we have characterized twenty Huntingtin 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 issues 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/13-922/v2UniProt ID P42858 HTT Huntingtin antibody characterization antibody validation western bloteng |
| spellingShingle | Rachel J. Harding Carl Laflamme Sara González Bolívar Aled M. Edwards Kathleen Southern Rebeka Fanti Riham Ayoubi Charles Alende Maryam Fotouhi Renu Chandrasekaran A guide to selecting high-performing antibodies for Huntingtin (UniProt ID: P42858) for use in western blot, immunoprecipitation, and immunofluorescence [version 2; peer review: 2 approved] F1000Research UniProt ID P42858 HTT Huntingtin antibody characterization antibody validation western blot eng |
| title | A guide to selecting high-performing antibodies for Huntingtin (UniProt ID: P42858) for use in western blot, immunoprecipitation, and immunofluorescence [version 2; peer review: 2 approved] |
| title_full | A guide to selecting high-performing antibodies for Huntingtin (UniProt ID: P42858) for use in western blot, immunoprecipitation, and immunofluorescence [version 2; peer review: 2 approved] |
| title_fullStr | A guide to selecting high-performing antibodies for Huntingtin (UniProt ID: P42858) 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 Huntingtin (UniProt ID: P42858) for use in western blot, immunoprecipitation, and immunofluorescence [version 2; peer review: 2 approved] |
| title_short | A guide to selecting high-performing antibodies for Huntingtin (UniProt ID: P42858) for use in western blot, immunoprecipitation, and immunofluorescence [version 2; peer review: 2 approved] |
| title_sort | guide to selecting high performing antibodies for huntingtin uniprot id p42858 for use in western blot immunoprecipitation and immunofluorescence version 2 peer review 2 approved |
| topic | UniProt ID P42858 HTT Huntingtin antibody characterization antibody validation western blot eng |
| url | https://f1000research.com/articles/13-922/v2 |
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