Serine/threonine-protein kinase Nek1, Thermo Fisher Scientific, PA5-78074

Our team has carried out deep research in the published scientific literature to identify papers and reports using antibodies directed against human Serine/threonine-protein kinase Nek1 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 6 commercial research antibodies against human Serine/threonine-protein kinase Nek1 encoded by NEK1

The collection of antibodies was tested using HeLa WT and NEK1 KO cells in the applications WB (WT and KO cell lysates/100 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) PA5-78074 detected a band of appr. 145 kDa in WT cell lysate with no corresponding band observed in KO cell lysate. Furthermore, a faint band of lower molecular weight was observed in WT cell lysate. A band was detected following IP (1 µg Ab used). This band was more intense than corresponding band in input extract (10 % of starting material), and only a faint band was observed in immunodepleted extract (10% of total unbound fraction) showing that the antibody does deplete the antigen from the cell lysate. In IF (1:300 ab dilution), a more intense staining of WT cells was observed compared to KO cell staining, showing some background.

We highly encourage other scientists to share their observations.