ab24859 staining TIE2 in Human spleen by Immunohistochemistry (Frozen sections).

Immunohistochemical analysis of Human brain tissue, staining TIE2 with ab24859.

Tissue was fixed with paraformaldehyde, permeabilized with 0.25 Triton X-100 and blocked with 2.5% BSA for 30 minutes at 25°C. Samples were incubated with primary antibody (1/200 in 2.5% horse serum) for 18 hours at 4°C. An HRP-conjugated horse anti-rabbit polyclonal IgG was used as the secondary antibody.

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Overlay histogram showing JEG-3 cells stained with ab24859 (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 (ab24859, 2µg/1x10⁶ cells) 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 JEG-3 cells fixed with 4% paraformaldehyde (10 min) used under the same conditions.

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

All lanes : Anti-TIE2 antibody [Cl. 16] (ab24859)

Lane 1 : HUVECs left untreated
Lane 2 : HUVECs stimulated for 3 hours with PMA at 25 ng/ml
Lane 3 : HUVECs stimulated for 6 hours with PMA at 25 ng/ml
Lane 4 : HUVECs stimulated for 9 hours with PMA at 25 ng/ml
Lane 5 : HUVECs stimulated for 24 hours with PMA at 25 ng/ml

Predicted band size: 126 kDa



Samples were immunoprecipitated with another TIE2 antibody.