DOI: 10.12688/f1000research.185471.1 ISSN: 2046-1402

A guide to selecting high-performing antibodies for PKN2 (UniProt ID: Q16513) for use in western blot and flow cytometry

Michael Biddle, Jemma Cooper, Carolyn Jones, Katie Dixon, Harvinder Virk
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Idiopathic pulmonary fibrosis is a chronic, progressive interstitial lung disease characterised by fibrotic remodelling of the distal alveolar interstitium and irreversible loss of lung function. Rapid decline in forced vital capacity has been significantly associated with rs115982800, a single nucleotide polymorphism in PKN2. Identifying high-quality research antibodies is therefore essential to enable robust investigation of PKN2 biology and its translational potential. Here, we systematically evaluated eleven commercial antibodies for western blot and flow cytometry using a standardised knockout-validation approach in human HCT 116 cells, comparing readouts in PKN2 knockout lines with isogenic parental controls. These experiments are part of a broader collaborative initiative addressing antibody reproducibility by characterising commercial antibodies for human proteins and making the results openly available. While antibody performance and protocol conditions may vary between laboratories, this study provides a resource to guide selection of the most suitable reagents for investigations of PKN2 in health and disease.

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