All lanes : Anti-Transglutaminase 2 antibody [CUB 7402] (ab2386) at 1 µg/ml

Lane 1 : Wild-type A549 whole cell lysate
Lane 2 : TGM2 knockout A549 whole cell lysate
Lane 3 : HeLa whole cell lysate
Lane 4 : Huvec whole cell lysate

Lysates/proteins at 20 µg per lane.

Predicted band size: 77 kDa
Observed band size: 77 kDa

Lanes 1 - 4: Merged signal (red and green). Green - ab2386 observed at 77 kDa. Red - loading control, ab181602, observed at 37 kDa.

ab2386 was shown to recognize  in wild-type A549 cells as signal was lost at the expected MW in TGM2 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and TGM2 knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% Milk. Ab2386 and ab181602 (Rabbit anti-GAPDH loading control) were incubated overnight at 4°C at 1 ug/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye^® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preabsorbed ab216777 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

Immunohistochemical analysis of breast carcinoma tissue labeling Transglutaminase 2 with ab2386. Tissue was fixed with paraformaldehyde; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/50) for 30 minutes at 22°C.

Overlay histogram showing HCT116 cells stained with ab2386 (red line). The cells were fixed with methanol (5 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab2386, 1/10 dilution) for 30 min at 22°C. The secondary antibody used was DyLight^® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a decreased signal in HCT116 cells fixed with 4% paraformaldehyde (10 min) used under the same conditions.

Immunohistochemical analysis of (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections) rat lung tissue labeling Transglutaminase 2 with ab2386. Tissue was fixed with formaldehyde and blocked with 1% casein in PBS for 15 minutes at room temperature; antigen retrieval was by heat mediation in a citrate buffer, pH 6. Samples were incubated with primary antibody (1/200 in TBS + BSA) for 20 minutes. ab98782 was used as the secondary antibody at a dilution of 1/400.

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