Millipore ABD51

Disclaimer: The fact that this page reports negative results with the given antibody does not mean that the antibody does not work – it just means that our laboratory did not succeed, using our typical experimental protocol, to find the searched protein by immunoblotting in the tissue examined. Modificiation of the protocol, or the selection of other tissue, might still provide excellent results.

The Millipore ABD51 has been reported to detect the bone morphogenetic protein receptor ALK3 in mouse liver lysates. We attempted to detect ALK3 in the 3000 g fraction of mouse liver homogenate, but the results are controversial. The antibody detects nice strong bands at approximately the correct position (slightly below 75 kDa); however, the bands do not respond at all to PNGase F treatment, which is in contrast to the one glycosylation reported by Uniprot database. In our 2018 paper we therefore conclude that this antibody is not sufficiently validated for in vivo use (Frýdlová J et al., Am J Physiol Gastrointest Liver Physiol. 2018 Oct 1;315(4):G560-G568). Moreover, we also had the opportunity to test this antibody on liver samples of mice with conditionally deleted Alk3 gene, there was no difference between wild type mice and Alk3-/- mice, the bands remained in place. Therefore, we are unable to confirm that the antibody indeed detects its target in the liver 3000 g homogenate fraction, which is nicely enriched in other hepatocyte plasma membrane proteins such as hemojuvelin or TMPRSS6. At present, we therefore do not regard this antibody as suitable for in vivo use.