alpha-1-antitrypsin, Sigma-Aldrich, A0409

Our team has carried out deep research in the published scientific literature to identify papers and reports using antibodies directed against human Alpha-1-antitrypsin (A1AT) 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 18 commercial research antibodies against human Alpha-1-antitrypsin (A1AT) encoded by SERPINA1

The collection of antibodies was tested using HepG2 WT and SERPINA1 KO cells in the applications WB (WT and KO culture media and cell lysate/30 µg protein), IP (WT culture medium) and flow cytometry (KO and WT cells) according to YCharOS standard protocols (see reference).

In WB (1:15000 ab dilution) A0409 did not detect a band specific to WT culture medium. A faint band of appr. 60 kDa was specific to WT cell lysate, however, a more intense band of higher molecular weight was observed in both genotypes. A band was detected following IP (2 µg Ab used), this band had same intensity compared to bands in input extract (4% of starting material) and immunodepleted extract (4% of total unbound fraction), showing that the antibody does not deplete the antigen from cell lysate. Flow cytometry using A0409 (1:1000 ab dilution) showed a right-shifted fluorescence intensity distribution in both WT and KO cells with minimal gap between the two populations, suggesting background staining in KO cells.