TGM2, Abcam, ab2386

I am sharing a pAbmAbs generated literature review of KO validated antibodies. Here citing Fell et al. Amino Acids 2021. Fell et al. validate this antibody for detection of endogenous human Transglutaminase 2 encoded by TG2 by Immunostaining of WT and TG2 KO idiopathic pulmonary fibrosis (IPF) fibroblasts. Faint staining was observed in TG2 KO compared to WT cells showing clear staining (Figure 6A). Based on the available data, we have assigned a four-star rating but we highly encourage other scientists to share their observations.

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 (1:1000 antibody dilution), ab2386 detected a band at the predicted size (~77 kDa) in WT A549 cell lysate, with no corresponding band in TGM2 KO lysate. Additionally, several weak bands were observed including a prominent band of lower molecular weight in the TGM2 KO lysate.

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 (1:1000 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.