Annexin A11, Abcam, ab236599

Our team has carried out deep research in the published scientific literature to identify papers and reports using antibodies directed against human Annexin A11 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 15 commercial research antibodies against human Annexin A11 encoded by ANAX11.

The collection of antibodies was tested using HeLa WT and ANAX11 KO cells in the applications WB (WT and KO cell lysates/50 µg protein), IP (WT cell lysate) and IF (WT and KO cells) according to YCharOS standard protocols (see reference). Western blot was furthermore carried out in HEK293T-, HAP1-, MDA-MB-231 pWPLXd (GFP positive)-, HCT 116- and hTERT-RPE1 WT cells lysates for verification of ab236599 antibody.

In WB (1:1000 Ab dilution) ab236599 detected a single band of appr. 54 kDa in WT lysate, with no band observed in KO lysate. Comparison of Annexin A11 protein level in different WT cell lines and HeLa KO showed a clear band for each cell lines and no band for HeLa KO by Western blotting (1:1000 Ab dilution). Only a weaker band was detected following IP (1 µg Ab used). However, the immunoprecipitated band was less intense than the corresponding band in input extract (10% of starting material) and immunodepleted extract (10% of total unbound fraction), showing that the antibody does not efficiently immunoprecipitate the antigen. In IF (1:400 Ab dilution), using PFA for fixation, identical staining patterns were observed in both KO and WT cells. In IF using Methanol for fixation a weak staining of WT cells was observed compared to KO cells. We highly encourage other scientists to share their observations