Western blot analysis of human Cervical cancer cell line (HeLa) lysate showing detection of 52.9 kDa ATG4D protein using Anti-ATG4D Antibody (A304858) at 1:1,000 for 1 hour at room temperature. Lane 1: Molecular Weight Ladder (MW). Lane 2: human HeLa cell lysates. Load: 15 µg. Block: 5% Skim Milk in 1X TBST. The secondary antibody used was Goat Anti-Rabbit HRP at 1:2000 for 60 minutes at room temperature. Color Development: ECL solution for 6 min in room temperature. Predicted/Observed Size: 52.9 kDa. Other Band(s): ~45 and 48 kDa.
Immunocytochemistry/Immunofluorescence analysis of human colon carcinoma cell line (RKO), fixed in 4% formaldehyde for 15 min at room temperature, using Anti-ATG4D Antibody (A304858), at 1:100 for 60 minutes at room temperature. The secondary antibody used was Goat Anti-Rabbit ATTO 488 at 1:100 for 60 minutes at room temperature. Counterstain: Phalloidin Texas Red F-Actin stain; DAPI (blue) nuclear stain at 1:1000, 1:5,000 for 60 minutes at room temperature, 5 minutes at room temperature. Localization: Cytoplasm. Magnification: 60X.(A) DAPI nuclear stain. (B) Phalloidin Texas Red F-Actin stain. (C) ATG4D Antibody. (D) Composite.
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