Mouse monoclonal [4F1] antibody to alpha Synuclein.
Specificity
This antibody binds multiple linear epitopes within human alpha synuclein: 21-70 aa, 61-90 aa, and 101-140 aa, determined using Dot blot and ELISA with corresponding peptides. Band corresponding to Alpha synuclein. Band at ~40 kDa is a trimer.
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 - Anti-alpha Synuclein Antibody [4F1] (A304962)
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.
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