Human/Mouse/Rat Transglutaminase 2/TGM2 Antibody

Detection of Human, Mouse, and Rat Transglutaminase 2/TGM2 by Western Blot. Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line, HepG2 human hepatocellular carcinoma cell line, mouse liver tissue, and rat liver tissue. PVDF membrane was probed with 0.5 µg/mL of Sheep Anti-Human/Mouse/Rat Transglutaminase 2/TGM2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4376) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (HAF016). A specific band was detected for Transglutaminase 2/TGM2 at approximately 85 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of Human Transglutaminase 2/TGM2 by Simple Western^TM. Simple Western lane view shows lysates of HeLa human cervical epithelial carcinoma cell line loaded at 0.2 mg/mL. A specific band was detected for Transglutaminase 2/TGM2 at approximately 82 kDa (as indicated) using 5 µg/mL of Sheep Anti-Human/Mouse/Rat Transglutaminase 2/TGM2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4376) followed by 1:50 dilution of HRP-conjugated Anti-Sheep IgG Secondary Antibody (HAF016). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Transglutaminase 2/TGM2 Specificity is Shown by Immunocytochemistry in Knockout Cell Line. A549 WT and Transglutaminase 2/TGM2 KO cells were labelled with a green or a far-red fluorescent dye, respectively. Cells were stained with Sheep Anti-Human/Mouse/Rat Transglutaminase 2/TGM2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4376) followed by incubation with a donkey anti-sheep Alexa-fluor 555 coupled secondary antibody (upper panel). DAPI-only counterstained cells shown on a lower panel. Acquisition of the blue (nucleus-DAPI), green (identification of WT cells), red (antibody staining) and far-red (identification of KO cells) channels was performed. Representative images of the blue and red (grayscale) channels are shown. WT and KO cells are outlined with green and magenta dashed line, respectively. Primary antibody concentration used: 0.2 µg/mL. Image, protocol and testing courtesy of YCharOS Inc. (ycharos.com).

Western Blot Shows Human Transglutaminase 2/TGM2 Specificity Using Knockout Cell Line. Western blot shows A549 cell line and Transglutaminase 2/TGM2 knockout A549 cell line (KO). A549 WT and Transglutaminase 2/TGM2 KO were cultured in serum free media, and 40 μg of protein from concentrated culture media were processed for western blot. Nitrocellulose membrane was probed with 0.5 µg/mL of Sheep Anti-Human/Mouse/Rat Transglutaminase 2/TGM2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4376) followed by HRP-conjugated donkey anti-sheep IgG Secondary Antibody. A specific band was detected for Transglutaminase 2/TGM2 at approximately 75 kDa (as indicated) in the parental A549 cell line, but is not detectable in knockout A549 cell line. The Ponceau stained transfer of the blot is shown. This experiment was conducted under reducing conditions. Image, protocol, and testing courtesy of YCharOS Inc. See ycharos.com for additional details.

Western Blot Shows Human Transglutaminase 2/TGM2 Specificity Using Knockout Cell Line. Western blot shows lysates of A549 cell line and Transglutaminase 2/TGM2 knockout A549 cell line (KO). Nitrocellulose membrane was probed with 0.5 µg/mL of Sheep Anti-Human/Mouse/Rat Transglutaminase 2/TGM2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4376) followed by HRP-conjugated donkey anti-sheep IgG Secondary Antibody. A specific band was detected for Transglutaminase 2/TGM2 at approximately 75 kDa (as indicated) in the parental A549 cell line, but is not detectable in knockout A549 cell line. The Ponceau stained transfer of the blot is shown. This experiment was conducted under reducing conditions. Image, protocol, and testing courtesy of YCharOS Inc. See ycharos.com for additional details.

Detection of Transglutaminase 2/TGM2 by Immunoprecipitation. PMA-treated A549 lysates were prepared and immunoprecipitation was performed using 2.0 µg of Sheep Anti-Human/Mouse/Rat Transglutaminase 2/TGM2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4376) pre-coupled to Dynabeads protein G. Immunoprecipitated Transglutaminase 2/TGM2 was detected with Rabbit Anti-Transglutaminase 2/TGM2 Antibody.The Ponceau stained transfer of the blot is shown. SM=4% starting material; UB=4% unbound fraction; IP=immunoprecipitate; HC=antibody heavy chain. Image, protocol and testing courtesy of YCharOS Inc. (ycharos.com).