Anti-O-GalNAC Antibody [9H6] (A304788)

Immunohistochemistry analysis of mouse colon, fixed in formalin. The Primary Antibody used was Anti-O-GalNAC Antibody [9H6] (A304788) at 1:25 for 1 hour at room temperature. The secondary antibody used was Goat Anti-Mouse IgG: Alexa Fluor 488. Counterstain: Actin-binding Phalloidin-Alexa Fluor 633; DAPI (blue) nuclear stain. Magnification: 63X.(A) DAPI (blue) nuclear stain. (B) Citrulline Antibody (C) Composite.

Western blot analysis of GlcNAc-BSA Conjugate showing detection of 67 kDa O-GlcNAc protein using Anti-O-GalNAC Antibody [9H6] (A304788) at 1:1,000 for 2 hours at room temperature. Lane 1: Molecular Weight Ladder (MW). Lane 2: BSA. Lane 3: GlcNAc-BSA. Lane 4: GalNAc-BSA. Lane 5: Galactose-BSA. Lane 6: Glucose-BSA. Lane 7: Citrulline-BSA. Load: 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-O-GalNAC Antibody [9H6] (A304788), 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) DAPI (blue) nuclear stain. (B) Phalloidin Alexa Fluor 633 F-Actin stain. (C) O-GlcNAc Antibody (D) Composite.