Beta-Dystroglycan1, Developmental Studies Hybridoma Bank, MANDAG2(7D11)

Our team has carried out deep research in the published scientific literature to identify papers and reports using antibodies directed against human Dystroglycan 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 Dystroglycan 1encoded by DAG1. Dystroglycan 1 is cleaved into the following 2 chains: Alpha-dystroglycan and Beta-dystroglycan. The antibodies tested has specificity for either one of the chains or both.

The collection of antibodies against Beta-dystroglycan was tested using A431WT and DAG1 KO cells in the applications WB (WT and KO cell lysates/90 µg protein) and IF (WT and KO cells), according to YCharOS standard protocols (see reference).

The collection of antibodies against Alfa-dystroglycan was tested using HAP1 WT and DAG1 KO cells in the applications WB (WT and KO cell lysates/100 µg protein) and IP (WT cell lysate), according to YCharOS standard protocols (see reference).

MANDAG2 (7D11) has specificity for Beta-Dystroglycan. In WB (1:500 Ab dilution) MANDAG2 (7D11) detected a band of appr. 130 kDa in A431 WT cell lysate with no corresponding band observed in A431 KO cell lysate. Multiple bands of lower molecular weight were observed for both genotypes. A band was also detected following IP (2 µg Ab used) from A431 WT cells. This band was more intense than the corresponding band in input extract (10% of starting material). Band was not detected in immunodepleted extract (10% of total unbound fraction), showing that the antibody does completely deplete the antigen from the cell lysate. In IF (1:50 Ab dilution), comparable staining intensities were observed in both KO and WT cells.

We highly encourage other scientists to share their observations.