Huntingtin, Cell Signaling Technology, 5656

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) 5656 detected a band of appr. 350 kDa in WT cell lysate with no corresponding band in KO cell lysate; additionally, a strong band of lower molecular weight was also detected in WT cell lysate. An intense band was detected following IP (2 µg Ab used), this band was markedly more intense than the corresponding band in input extract (4% of starting material). Only a faint band was detected in immunodepleted extract (4% of total unbound fraction). In IF (1:100 Ab dilution) a clear intense staining was observed in WT cells with no background staining in KO cells.

We highly encourage other scientists to share their observations.