CD44, Abcam, ab189524

Our team has carried out deep research in the published scientific literature to identify papers and reports using antibodies directed against human CD44 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 32 commercial research antibodies against human CD44.

The collection of antibodies was tested using MDA-MB-231 WT and CD44 KO cells in the applications WB (WT and KO cell lysates/30 µg protein), IP (WT cell lysate), IF (WT and KO cells) and FC (WT and KO cells), furthermore, WB (WT and KO cell lysates/30 µg protein) for ab189524 was carried out using HAP1 WT and CD44 KO, according to YCharOS standard protocols (see reference).

Three applications of WB were done with ab189524. In the first WB (1:1000 Ab dilution) ab189524 detected a single band of appr. 81 kDa in MDA-MB-231 WT lysate, with no corresponding band observed in MDA-MB-231 KO lysate. Lysates of HAP1 WT and CD44 KO were prepared and applied for WB (1:1000 Ab dilution), detecting a single strong band of appr. 81 kDa in HAP1 WT, with no corresponding band in CD44 KO.
Furthermore, protein extracts were prepared in RIPA buffer from MDA-MB-231 WT and CD44 KO as well as in IP buffer from MDA-MB-231 WT and applied for WB (1:3000 Ab dilution), where ab189524 detected a band of appr. 81 kDa in WT lysates with no corresponding band in KO lysates.
No band was detected following IP (2 µg Ab used). In IF (1:250 Ab dilution), a clear intense staining was observed in WT cells with no background staining in KO cells. Cell surface and intracellular flow cytometry (1 µg/mL antibody) showed a clear fluorescence shift in the WT cell population compared with the KO cells, indicating specific binding of the antibody to the target protein.
We highly encourage other scientists to share their observations.