All lanes : Anti-BIRC6/APOLLON antibody (ab19609) at 1/10000 dilution

Lane 1 : HeLa whole cell lysate at 20 µg
Lane 2 : HeLa whole cell lysate at 50 µg
Lane 3 : HeLa S100 extract at 20 µg
Lane 4 : HeLa S100 extract at 50 µg

Predicted band size: 528 kDa

Detection of Human BIRC6/APOLLON by western blot.
Lane 1:  20 µg of HeLa whole cell lysate.
Lane 2:  50 µg of HeLa whole cell lysate.
Lane 3:  20 µg of HeLa S100 extract.
Lane 4:  50 µg of HeLa S100 extract.

Affinity purified anti-BIRC antibody (ab19609) was used at a 1/10,000 dilution.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue labelling BIRC6/APOLLON with ab19609 at 1/5000 (0.2µg/ml). Detection: DAB.

Detection of human BIRC6/APOLLON by Immunoprecipitation. Affinity purified ab19609 used at 2mcg/mg lysate (whole cell lysate (10mg) from HeLa cells. The immunoprecipitate was resolved by SDS-PAGE and stained with Coomassie Blue. The band that is BIRC6/APOLLON was confirmed by mass spectrometry.

ICC/IF image of ab19609 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab19609, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.