Netrin-1, Abcam, ab126729

Our team has carried out deep research in the published scientific literature to identify papers and reports using antibodies directed against human Netrin-1 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 14 commercial research antibodies against human Netrin-1 encoded by NTN1.

The antibody collection was evaluated using MCF7 WT and NTN1 KO cells in Western blot (WB; WT and KO cell culture medium, 10 µg protein) and immunoprecipitation (IP; WT culture medium), following YCharOS standard protocols (see reference). In addition, ab126729 was tested by WB (10 µg protein) across multiple cell lines, including WT and NTN1 KO MCF7, WT and NTN1 KO U-2 OS, as well as WT U-87 MG, HeLa, HT-1080, SK-N-FI, A549, A-431, 293T, and OVCAR3 cells.

In WB (1:2000 ab dilution) on MCF7 WT and NTN1 KO culture medium, ab126729 detected a band of appr. 75 kDa in WT culture medium, with no corresponding band observed in KO culture medium. In Western blot (1:2000 antibody dilution) across multiple cell lines, a band at approximately 75 kDa was observed in MCF7 WT, U-2 OS WT, A-431-, 293T-, and OVCAR3 WT on culture medium, indicating Netrin-1 expression in these cells. In contrast, only faint bands were detected in cell lysates. A band was also detected following IP (2 µg Ab used). This band was markedly more intense than the corresponding band in input extract (10% of starting material). No band was detected in immunodepleted extract (10% of total unbound fraction), showing that the antibody does completely deplete the antigen from the cell lysate.

We highly encourage other scientists to share their observations.