A guide to selecting high-performing antibodies for PLC-gamma-2 for use in Western Blot, immunoprecipitation and immunofluorescence
Open accessAbstract
Phosphatidylinositol-specific phospholipase C gamma 2 (PLC-gamma-2) is an enzyme that regulates the function of immune cells. PLC-gamma-2 has been implicated in neurodegenerative and autoimmune disorders, yet investigation of this protein has been limited by a lack of independently characterized antibodies. Here we have characterized eleven PLC-gamma-2 commercial antibodies for use in 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.
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PLCG2 antibody selection was validated by YCharOS.
“PLCG2 antibody selection was validated by YCharOS. 34”
From Alzheimer's disease–associated PLCG2 variants alter microglial state and function in human induced pluripotent stem cell–derived microglia‐like cells (2025) · cites this paper as [34]
The Introduction and Results and discussion (pages not provided) describe testing eleven PLC-gamma-2 antibodies and identifying several that performed well under the study’s conditions. This supports using the characterization data to guide antibody selection, but the authors explicitly say they do not offer antibody recommendations; the paper does not establish that a particular selection was validated by YCharOS. (LLM, conf=0.84)
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“After 34 After overnight incubation, coverslips were washed three times in PBST and incubated for 1 h at RT with the appropriate species-specific secondary antibodies conjugated with AlexaFluor in 5% BSA in PBST (1:1000, Invitrogen).”
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- Authors
- Ruíz Moleón, V., Fotouhi, M., Alende, C., Ayoubi, R., Bedford, L. M., Southern, K., Richardson, T. I., Laflamme, C., NeuroSGC/YCharOS/EDDU collaborative group, & ABIF consortium
- Affiliations
- Montreal Neurological Institute and Hospital (CA)
- Indiana University Indianapolis (US)
- Corresponding author
- Laflamme, Carl
- ORCID
- Venue
- F1000Research
- Publisher
- F1000 Research Ltd
- Published
- 2024-01-18
- Volume
- 13
- Pages
- 77
- ISSN
- 2046-1402 (ISSN-L)
- PMC
- PMC11489847
- OpenAlex
- W4390986454
- Cited by
- 1
- References
- 36
- Citations by year
- 2025: 1
- FWCI
- 0.25
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- F1000Research (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- IUScholarWorks (Indiana University) (submittedVersion, cc-by)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Funders
- Mitacs
- Canadian Institutes of Health Research Foundation (FDN154305)
- Genome Québec
- Genome Canada
- Ontario Genomics (OGI-210)
- National Institute on Aging (U54AG065187)
- Indiana University School of Medicine Purdue University TaRget Enablement to Accelerate Therapy Development for Alzheimer’s Disease
- Topics
- Health Sciences › Medicine › Radiology, Nuclear Medicine and Imaging › Monoclonal and Polyclonal Antibodies Research
- Life Sciences › Immunology and Microbiology › Immunology › T-cell and B-cell Immunology
- Physical Sciences › Chemistry › Spectroscopy › Advanced Proteomics Techniques and Applications
- Keywords
- commercial antibodies, antibody characterization, antibody reproducibility, knockout cell lines, Western blot, immunoprecipitation, immunofluorescence, neurodegenerative diseases, autoimmune disease
- MeSH terms
- Antibodies, Fluorescent Antibody Technique, Humans, Blotting, Western, Immunoprecipitation, Phospholipase C gamma
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