Anti-Nitrotryptophan Antibody [4F8] (A304724)

Immunocytochemistry/Immunofluorescence analysis of human embryonic kidney epithelial cell line (HEK293), fixed in 5% formaldehyde for 5 minutes, using Anti-Nitrotryptophan Antibody [4F8] (A304724), 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. Localization: Cytoplasmic. Magnification: 20X (2X Zoom).(A) DAPI (blue) nuclear stain. (B) Phalloidin Alexa Fluor 633 F-Actin stain. (C) Nitrotryptophan Antibody. (D) Composite.

Western blot analysis of 6-Nitrotryptophan-BSA Conjugate showing detection of 67 kDa Nitrotryptophan protein using Anti-Nitrotryptophan Antibody [4F8] (A304724) at 1:1,000 for 2 hours at room temperature. Lane 1: Molecular Weight Ladder (MW). Lane 2: BSA (0.5 µg). Lane 3: BSA (1 µg). Lane 4: 6-Nitrotryptophan-BSA (0.5 µg). Lane 5: 6-Nitrotryptophan-BSA (1 µg). Lane 6: 7-Ketocholesterol-BSA (0.5 µg). Lane 7: 7-Ketocholesterol-BSA (1 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: Luminol for 1 min. Predicted/Observed Size: 67 kDa.

Immunohistochemistry analysis of human thyroid cancer. The Primary Antibody used was Anti-Nitrotryptophan Antibody [4F8] (A304724) at 1:100 for Overnight at 4°C, then 30 minutes at 37°C. The secondary antibody used was Goat Anti-Mouse IgG (H+L): FITC for 45 minutes at 37°C. Counterstain: DAPI for 3 min at room temperature. Magnification: 5X.