Immunohistochemical analysis of human stomach tissue labeling Mucin 5AC with ab3649 at 2 µg/ml.

Overlay histogram showing A549 (Human lung carcinoma cell line) cells stained with ab3649 (red line). The cells were fixed with 4% paraformaldehyde (10 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 (ab3649, 1µg/1x10⁶ cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. 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 positive signal in A549 cells fixed with 80% methanol (5 min) used under the same conditions.

Please note that Abcam do not have any data for use of this antibody on non-fixed cells. We welcome any customer feedback.

Immunohistochemical analysis of human gastric carcinoma tissue labeling Mucin 5AC with ab3649 at 2 µg/ml.

Immunohistochemical analysis of rat stomach tissue labeling Mucin 5AC with ab3649 at 2 µg/ml.

Immunohistochemical analysis of rat stomach tissue, staining Mucin 5AC with ab3649.

Antigen retrieval was performed via heat mediation in a citrate buffer. Samples were blocked before incubating with primary antibody (1/100) for 1 hour at room temperature. An HRP-conjugated anti-mouse IgG was used as the secondary antibody and staining was detected using DAB.