A guide to selecting high-performing antibodies for Synaptotagmin-1 (Uniprot ID P21579) for use in western blot, immunoprecipitation, immunofluorescence and flow cytometry

Author:

Biddle Michael S.ORCID,Alende CharlesORCID,Fotouhi Maryam,Jones Carolyn,Ayoubi Riham,Southern KathleenORCID,Laflamme CarlORCID,Virk HarvinderORCID, ,

Abstract

Synaptotagmin-1 is a synaptic vesicle transmembrane protein that senses calcium influx via its tandem C2-domains, triggering synchronous neurotransmitter release. Disruption to SYT1 is associated with neurodevelopmental disorders, highlighting the importance of identifying high-quality research reagents to enhance understanding of Synaptotagmin-1 in health and disease. Here we have characterized thirteen Synaptotagmin-1 commercial antibodies for western blot, immunoprecipitation, immunofluorescence and flow cytometry 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

National Institute on Aging

Accelerating Medicines Partnership Program for Alzheimers disease

Ontario Genomics

Genome Canada

Genome Quebec

Publisher

F1000 Research Ltd

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