Huntingtin, Thermo Fisher Scientific, MA3-040

Our team has carried out deep research in the published scientific literature to identify papers and reports using antibodies directed against human Huntingtin and followed up by extraction and evaluation of the available data to assess its performance according to pAbmAbs rating criteria.

YCharOS is here conducting a large head-to-head study comparing 20 commercial research antibodies against human Huntingtin encoded by HTT.

The collection of antibodies was tested using DMS35 WT and HTT KO cells in the applications WB (WT and KO cell lysates/30 µg protein), IP (WT cell lysate) and IF (WT and KO cells) according to YCharOS standard protocols (see reference).

In WB (1:1000 Ab dilution) MA3-040 detected a band of appr. 350 kDa in WT cell lysate with no corresponding band in KO cell lysate. Furthermore, a faint band of lower molecular weight was observed in WT cell lysate. A double band corresponding to the target protein was detected in the IP sample following immunoprecipitation using 2 μg antibody. The signal appears stronger than in the input lane (4% of starting material), consistent with enrichment of the target protein during IP. A band was still present in the immunodepleted (unbound) fraction (4%), indicating that the antibody does not fully deplete the antigen from the lysate. In IF (1:1000 Ab dilution), comparable staining intensity was observed in both genotypes.

We highly encourage other scientists to share their observations.