Alpha Diagnostics International HFE11-A

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.

 

A puzzling antibody! In experiments with HFE KO mice the signal in liver plasma-membrane enriched fraction was much lower than in wild type mice, suggesting that the antibody successfully detects HFE. However, when we had the opportunity to test HFE content in liver samples from mice overexpressing HFE (AAV2/8 vector, samples generously provided by Dr. Andrea Steinbicker), we detected no extra signal at all. Therefore, our conclusion must be that the antibody does not reliably detect HFE (and all the nice blots we have done using this antibody can be thrown away…)

 

 

Liver samples from various mice (L1-L4) probed with the HFE antibody. HFE KO mouse liver shows almost no signal in the expected range – a (almost) conclusive evidence that the antibody really detects HFE.

 

And then, a disappointing result with livers from HFE overexpressing mice:

 

 

Since the Santa Cruz polyclonal antibody WAS able to see dramatic overexpression of HFE in the same samples of livers from mice injected with HFE AAV8, the current conclusion must be that this antibody can not be used for reliable in vivo HFE protein detection.