Anti-alpha Synuclein Antibody [4F1] (A304962)

Immunocytochemistry/Immunofluorescence analysis of rat primary hippocampal neurons treated with active alpha synuclein protein aggregate (SPR-322) at 4 µg/ml to induce fibrils, fixed in 4% paraformaldehyde, using Anti-alpha Synuclein Antibody [4F1] (A304962), at 1:200 for 24 hours at 4°C. The secondary antibody used was Goat Anti-Mouse Alexa Fluor 488 at 1:700 for 1 hour at room temperature. Counterstain: Guinea Pig Anti-NeuN (red) neuronal marker (Donkey Anti-Guinea Pig Alexa Fluor 647 1:700); DAPI (blue) nuclear stain at 1:6000, 1:3000 for 60 minutes at room temperature, 5 minutes at room temperature. Magnification: 20X.(A) DAPI (blue) nuclear stain. (B) NeuN neuronal marker (red). (C) Alpha Synuclein Antibody. (D) Composite.

Western blot analysis of Human, mouse, rat brain showing detection of 14 kDa Alpha Synuclein protein using Anti-alpha Synuclein Antibody [4F1] (A304962) at 1:1,000 for 2 hours at room temperature. Lane 1: Molecular Weight Ladder (MW). Lane 2: mouse brain cell lysate. Lane 3: rat brain cell lysate. Lane 4: human brain cell lysate. Load: 15 µg. Block: 5% Skim Milk in 1X TBST. The secondary antibody used was Goat Anti-Mouse HRP:IgG at 1:3000 for 1 hour at room temperature. Color Development: ECL solution (Super Signal West Pico) for 5 minutes in room temperature. Predicted/Observed Size: 14 kDa. Other Band(s): ~40 kDa (trimer).

Immunocytochemistry/Immunofluorescence analysis of human neuroblastoma cell line (SK-N-BE, fixed in 4% formaldehyde for 15 min at room temperature, using Anti-alpha Synuclein Antibody [4F1] (A304962), at 1:100 for 60 minutes at room temperature. The secondary antibody used was Goat Anti-Mouse ATTO 488 at 1:200 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: medium; Nucleus: strong. Magnification: 60X.(A) DAPI (blue) nuclear stain. (B) Phalloidin Texas Red F-Actin stain. (C) Alpha Synuclein Antibody. (D) Composite.