A guide to selecting high-performing antibodies for Huntingtin (UniProt ID: P42858) for use in western blot, immunoprecipitation, and immunofluorescence

Author:

Fanti Rebeka,Ayoubi Riham,Alende CharlesORCID,Fotouhi Maryam,González Bolívar SaraORCID,Chandrasekaran Renu,Southern KathleenORCID,Edwards Aled M.,Harding Rachel J.ORCID,Laflamme CarlORCID, ,

Abstract

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.

Funder

Mitacs

Genome Quebec

Ontario Genomics

Genome Canada

Publisher

F1000 Research Ltd

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