Immunohistochemistry analysis labeling human adrenal carcinoma tissue with ab240918 at a 1/100 dilution. Tissue samples were incubated with the primary antibody ab240918 at a 1/100 dilution overnight at 4^oC. A Goat anti-Mouse IgG (H&L):FITC secondary was used (45 min at 37^oC). Tissue samples were counterstained with DAPI (blue) for 3 minutes at room temperature. Magification 5X.

All lanes : Anti-Acrolein antibody [10A10] (ab240918) at 1/1000 dilution

Lane 1 : HeLa (human epithelial cell line from cervix adenocarcinoma) cell lysate
Lane 2 : Hydrogen peroxide-treated HeLa cell lysate

Lysates/proteins at 12 µg per lane.

Secondary
All lanes : HRP-linked goat anti-mouse IgG at 1/2000 dilution

Developed using the ECL technique.

Exposure time: 5 minutes

Block: 5% Skim Milk in TBST.

Primary incubation for 2 hours at RT.

Flow cytometric analysis of 90% methanol-fixed SH-SY5Y (human neuroblastoma cell line from bone marrow) cells labeling Acrolein using ab240918 at 1/50 dilution for 30 minutes on ice (solid line) compared to a non-specific IgG control (dashed line). Secondary antibody was a PE-conjugated goat anti-mouse IgG at 1/100 dilution for 20 minutes at room temperature. Cells were subject to oxidative stress by treating with 250 µM H₂O₂ for 24 hours.

All lanes : Anti-Acrolein antibody [10A10] (ab240918) at 1/1000 dilution

Lane 1 : Acrolein-BSA at 0.5 µg
Lane 2 : Acrolein-BSA at 2 µg
Lane 3 : BSA at 0.5 µg
Lane 4 : BSA at 2 µg

Secondary
All lanes : HRP-linked goat anti-mouse IgG at 1/2000 dilution

Developed using the ECL technique.

Exposure time: 5 minutes

Block: 5% Skim Milk in TBST.

Primary incubation for 2 hours at RT.