Anti-4-Hydroxynonenal Antibody [12F7] (A304722)

Western blot analysis of 4-hydroxy-nonenal-BSA Conjugate showing detection of 67 kDa 4-hydroxy-nonenal protein using Anti-4-Hydroxynonenal Antibody [12F7] (A304722) at 1:1,000 for 2 hours at room temperature. Lane 1: Molecular Weight Ladder (MW). Lane 2: BSA (0.5 µg). Lane 3: 4-hydroxyl nonenal-BSA (0.5 µg). Lane 4: 4-hydroxy nonenal-BSA (2.0 µg). Lane 5: 4-hydroxy-2-hexenal (0.5 µg). Lane 6: 4-hydroxy-2-hexenal (2.0 µg). Block: 5% Skim Milk in TBST. The secondary antibody used was Goat Anti-Mouse IgG: HRP at 1:2000 for 60 minutes at room temperature. Color Development: ECL solution for 5 minutes in room temperature. Predicted/Observed Size: 67 kDa.

Immunocytochemistry/Immunofluorescence analysis of human embryonic kidney epithelial cell line (HEK293), fixed in 5% formaldehyde for 5 minutes, using Anti-4-Hydroxynonenal Antibody [12F7] (A304722), at 1:50 for 30-60 min at room temperature. The secondary antibody used was Goat Anti-Mouse Alexa Fluor 488 at 1:1500 for 30-60 min at room temperature. Counterstain: Phalloidin Alexa Fluor 633 F-Actin stain; DAPI (blue) nuclear stain at 1:250, 1:50,000 for 30-60 min at room temperature. Magnification: 20X (2X Zoom). (A,C,E,G) Untreated. (B,D,F,H) Cells cultured overnight with 50 µM H2O2. (A,B) DAPI (blue) nuclear stain. (C,D) Phalloidin Alexa Fluor 633 F-Actin stain. (E,F) 4-Hydroxynonenal Antibody. (G,H) Composite.