Anti-PHD3 antibody

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-PHD3 antibody (AB30782)

ICC/IF image of ab30782 stained HepG2 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 ab30782 at 5μg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/1000 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.

• WB

Ap16142****

Western blot - Anti-PHD3 antibody (AB30782)

This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Bovine Serum Albumin before being incubated with ab30782 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.

All lanes:

Western blot - Anti-PHD3 antibody (ab30782) at 1 µg/mL

Lane 1:

Western blot - HeLa-Vehicle treated (Negative Control) Whole Cell Lysate (<a href='/en-us/products/cell-lysates/hela-vehicle-treated-negative-control-whole-cell-lysate-ab116321'>ab116321</a>) at 20 µg

Lane 2:

Hela-DFO treated (0.5mM, 24h) Whole Cell Lysate (<a href='/en-us/products/unavailable/hela-dfo-treated-05mm-24h-whole-cell-lysate-ab116322'>ab116322</a>) at 20 µg

Lane 3:

Human HeLa Nuclear DFO treated at 20 µg

Lane 4:

Human HeLa Cytoplasmic DFO treated at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/10000 dilution

Predicted band size: 27 kDa

Observed band size: 18 kDa,27 kDa

true

Exposure time: 20min

• WB

AbReview36191****

Western blot - Anti-PHD3 antibody (AB30782)

All lanes:

Western blot - Anti-PHD3 antibody (ab30782) at 1/1000 dilution

Lane 1:

Mouse embroyonic fibroblast cell lysate. Treated 21% O2 for 24 hours at 5 µg

Lane 2:

Mouse embroyonic fibroblast cell lysate. Treated 1% O2 for 24 hours at 5 µg

Secondary

All lanes:

Goat polyclonal anti-rabbit HRP-conjugate at 1/10000 dilution

Predicted band size: 27 kDa

true

Exposure time: 30s

This image is courtesy of an anonymous Abreview