pro-Neurotrophin 3, Alomone Labs, ANT-012

Our team has carried out deep research in the published scientific literature to identify papers and reports using ANT-012 and followed up by extraction and validation of the available data to assess its performance according to pAbmAbs rating criteria.
ANT-012 has been used in independent studies albeit from the same group (Journal of Neuroinflammation, 2013; PLOS ONE, 2014) to detect the precursor form of neurotrophin-3 (pro-NT3) in retinal tissue lysates by Western blot.
In Müller cell ablation models (J Neuroinflammation 2013), ANT-012 detected a clear upregulation of pro-NT3, with a distinct band visible in Western blot (Figure 3). Results were consistent and aligned with biological expectations (reduction in mature NT3, increase in pro-NT3).
In RCS rats treated with EPO (PLOS ONE 2014), ANT-012 was used to measure pro-NT3 expression changes. The antibody reliably showed that EPO treatment attenuated pro-NT3 upregulation during photoreceptor degeneration (Results, Fig. 8).
No evidence of off-target bands was reported; the antibody resolved pro-NT3 changes in line with physiological predictions, suggesting that the antibody is specific.
Limitations: No direct knockout or peptide competition controls were described; thus, specificity rests on expected expression patterns and consistency. Full gel images are not shown, and blots are quite narrow.
We highly encourage other scientists to share their observations.

References:
https://jneuroinflammation.biomedcentral.com/articles/10.1186/1742-2094-10-137#additional-information
https://journals.plos.org/plosone/article?id=10.1371%2Fjournal.pone.0104759

I have used it for IHF on brain sections of adult mice, with the dilution 1:100, it worked well, staining cytoplasm. However, I tried it for embryo sections and I do have some staining in cytoplasm, but not for individual cells, well delimitated, as it was with brain sections. Then, i’m not sure of its staining in early developmental stage.

We have used this antibody for WB detection of proNT3 in inner ear tissue from rat, as well as for detection of human recombinant proNT3. The antibody shows no cross-reactivity with proNGF and does not recognize the mature form of NT3. Please, see Eur J Neurosci. 2011 Feb;33(4):622-31 for further information.

The antibody appears to be good and specific for staining proNT3 in PFA-fixed cryosections of mouse embryos. In the developing CNS, only very distinct cell types are stained.

IF dilution 1:200