ICC/IF image of ab109088 stained HepG2 cells. The cells were 100% methanol fixed (5 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 ab109088 at 1/200 dilution overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) 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.
![All lanes : Anti-PHD2 / prolyl hydroxylase antibody [EPR3661] (ab109088) at 1/1000 dilutionLane 1 : SH-SY5Y cell lysate, treated with CoCl2Lane 2 : SH-SY5Y cell lysateLane 3 : A549 cell lysateLysates/proteins at 10 µg per lane.](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_4/10872_PHD2-prolyl-hydroxylase-Primary-antibodies-ab109088-1.jpg)
All lanes : Anti-PHD2 / prolyl hydroxylase antibody [EPR3661] (ab109088) at 1/1000 dilutionLane 1 : SH-SY5Y cell lysate, treated with CoCl2Lane 2 : SH-SY5Y cell lysateLane 3 : A549 cell lysateLysates/proteins at 10 µg per lane.
ab109088 at 1/50 dilution staining PHD2 / prolyl hydroxylase in Human kidney by Immunohistochemistry, Paraffin-embedded tissue
Overlay histogram showing SH-SY5Y cells stained with ab109088 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then 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 (ab109088, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in SH-SY5Y cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.