TGM2, R&D Systems, AF4376

Our team performed a comprehensive review of the scientific literature to identify studies using antibodies against human protein-glutamine gamma-glutamyltransferase 2 (TGM2), followed by extraction and evaluation of available data according to pAbmAbs rating criteria.

YCharOS conducted a systematic head-to-head comparison of 17 commercially available antibodies against TGM2 across multiple applications.

Antibodies were tested by WB using lysates (40 µg protein) from A549 WT (ab275463) and TGM2 KO (ab261876) cells. These cell lines were also used for IF. In addition, antibodies were evaluated by IP using A549 WT lysate (1 mg protein). All experiments were performed according to YCharOS standard protocols (see reference).

In WB (1:400 antibody dilution), AF4376 detected a clear band at the predicted size (~77 kDa) in WT A549 cell lysate, with no corresponding band in TGM2 KO lysate. Additionally, several weaker bands were observed in both cell lysates.

In IP, the antibody (2 µg) enriched TGM2 upon precipitation. However, target remains detectable in the unbound fraction indicating incomplete precipitation. TGM2 levels in the starting material, unbound fraction, and precipitate were assessed by WB using the TGM2-specific antibody A21184 from ABclonal (1:10.000).

In IF, the antibody (1:100 dilution) produced a TGM2-specific staining distributed throughout A549 WT cells, with no background signal in TGM2 KO cells.

We highly encourage other scientists to share their observations.