Santa Cruz sc-514405
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.
Remember how Santa Cruz marketing used to send us funny pictures stating that using polyclonals is like flying biplanes, while using monoclonals is like travelling in modern business jets?
Well, there ARE people who would definitely prefer a Stearman Kaydet over a Suchoj Superjet… While liver HFE definitely could be detected in the past using the sc-18810 polyclonal, the new sc-514405 monoclonal fails to detect anything resembling HFE in rat plasma membrane-enriched fraction, or even in samples from Hfe-overexpressing mice.
The blot on the left shows liver homogenate (in NP40-containing buffer) from control (C) and HFE-overexpressing (AAV2/8 virus) mice (HFE). Probing with the (now discontinued) 18810 polyclonal shows beautiful HFE bands (arrows). On the other hand, the 514405 monoclonal shows no trace of the overexpressed HFE protein. Blot B uses exactly the same samples as blot A. As both antibodies are reported to recognise a sequence within the internal part of the protein, the lack of signal in blot B indicates that something must be wrong…
The conclusion therefore is that the sc-514405 is not suitable for in vivo experiments. Interestingly, the antibody is sometimes used in high-impact publications (Liu J et al, HFE inhibits type I IFNs signaling by targeting the SQSTM1-mediated MAVS autophagic degradation. Autophagy. 2020 Aug 18:1-16).