Western blotting analysis using transglutaminase 2 antibody (Cat#63732). Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with transglutaminase 2 antibody (Cat#63732, 1:5,000) and HRP-conjugated goat anti-mouse secondary antibody (Cat#101, 1:20,000) respectively. Image was developed using FeQ™ ECL Substrate Kit (Cat#226).
Western blotting analysis using transglutaminase 2 antibody (Cat#63732). Transglutaminase 2 expression in wild-type (WT) and transglutaminase 2 (TGM2) shRNA knockdown (KD) HT-1080 cells with 20 μg of total cell lysates. β-Tubulin serves as a loading control. The blot was incubated with transglutaminase 2 antibody (Cat#63732, 1:5,000) and HRP-conjugated goat anti-mouse secondary antibody (Cat#101, 1:20,000) respectively. Image was developed using NaQ™ ECL Substrate Kit (Cat#716).
Immunohistochemistry was performed on paraffin-embedded human breast carcinoma using transglutaminase 2 antibody (Cat#63732, 1:200). Antigen retrieval was done in sodium citrate buffer (pH 6.0). DAB was used for detection, with hematoxylin counterstaining. Images were acquired using a Nikon Ci-L Plus microscope (40× objective). Scale bar: 25 μm.
Flow cytometric analysis of Transglutaminase 2 expression in HT-1080 cells using Transglutaminase 2 antibody (Cat#63732, 1:2,000). Fixative solution: 4% Paraformaldehyde in 1x PBS; Permeabilization solution: 0.5% Triton X-100 in 1x PBS. Green, isotype control; red, Transglutaminase 2.
Validation of Transglutaminase 2 knockdown using flow cytometry. Wild-type(WT, Blue) and knockdown(KD, Green) HT-1080 cells were stained with Transglutaminase 2 antibody (Cat#63732, 1:2,000) and analyzed using BD flow cytometer. Fixative solution: 4% Paraformaldehyde in 1x PBS; Permeabilization solution: 0.5% Triton X-100 in 1x PBS.
