Na+/Ca2+ exchange protein 3, Alomone Labs, ANX-013

We found one peer-reviewed publication specifically describing the validation of ANX-013 from Alomone Labs. Piccialli et al. (2021). “The Na+/Ca²⁺ Exchanger 3 Is Functionally Coupled With the NaV1.6 Voltage-Gated Channel and Promotes an Endoplasmic Reticulum Ca²⁺ Refilling in a Transgenic Model of Alzheimer’s Disease.” Frontiers in Pharmacology 12:775271. https://www.frontiersin.org/journals/pharmacology/articles/10.3389/fphar.2021.775271/full The NCX3 antibody from Alomone Labs was used for Western blotting, and was applied to nitrocellulose membranes at 1:1,000. Detection was by chemiluminescence, followed by film development and densitometric analysis. The samples were lysates from primary hippocampal neurons derived from wild-type and Tg2576 mice. Relevant validation evidence in the article: The strongest validation experiment was target-specific siRNA knockdown: The study compared NCX3 immunoreactivity in control neurons and neurons treated with NCX3-directed siRNA. The Western blot showed that NCX3 silencing reduced the NCX3 signal, while expression of the related exchanger isoforms NCX1 and NCX2 were not affected. The figure legend identifies this as a representative Western blot and quantification of NCX3, NCX1 and NCX2 after NCX3 silencing (Figure 2c). This constitutes: • Genetic validation: reduction of the antibody signal following NCX3 knockdown. • Isoform-selectivity support: the NCX3-directed intervention did not alter NCX1 or NCX2 expression. • Biological consistency: silencing NCX3 also prevented the increase in NCX-mediated current observed in Tg2576 neurons (Figure 2a-b). The antibody also detected an NCX3-related band at approximately 65 kDa, described as a truncated NCX3 form. This band was increased in Tg2576 neurons relative to wild-type neurons (Figure 2f-g). We have rated the antibody based on the available information and according to the pAbmAbs review criteria. We highly encourage other scientists to share their experience.