This antibody does not cross-react with Ataxin-1 (phosphor S751). This antibody exhibits minimal cross-reactivity with S751A mutant of Ataxin-1 by ELISA and immunofluorescence and is negative by immunoblot.
Figure 1: Western Blot - Anti-Ataxin 1 Antibody [S65-37] (A304806)
Western blot analysis of Monkey COS-1 cells transfected with Ataxin- 1 showing detection of ~85 kDa Ataxin 1 protein using Anti-Ataxin 1 Antibody [S65-37] (A304806) at 1:200 for 16 hours at 4°C. Lane 1: Molecular Weight Ladder. Lane 2: Monkey COS-1 cells transfected with Ataxin- 1. Load: 15 µg. Block: 2% BSA and 2% Skim Milk in 1X TBST. The secondary antibody used was Goat Anti-Mouse IgG: HRP at 1:1,000 for 1 hour room temperature. Color Development: ECL solution for 6 min in room temperature. Predicted/Observed Size: ~85 kDa.
Immunocytochemistry/Immunofluorescence analysis of human neuroblastoma cell line (SK-N-BE, fixed in 4% formaldehyde for 15 min at room temperature, using Anti-Ataxin 1 Antibody [S65-37] (A304806), at 1:100 for 60 minutes at room temperature. The secondary antibody used was Goat Anti-Mouse 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 room temperature, 5 minutes room temperature. Localization: Cytoplasm, Nucleus. Magnification: 60X.(A) DAPI (blue) nuclear stain. (B) Phalloidin Texas Red F-Actin stain. (C) Ataxin 1 Antibody. (D) Composite.
Immunocytochemistry/Immunofluorescence analysis of human neuroblastoma cells (SH-SY5Y), fixed in 4% PFA for 15 min, using Anti-Ataxin 1 Antibody [S65-37] (A304806), at 1:50 for overnight at 4°C with slow rocking. The secondary antibody used was AlexaFluor 488 at 1:1,000 for 1 hour at room temperature. Counterstain: Phalloidin-iFluor 647 (red) F-Actin stain; Hoechst (blue) nuclear stain at 1:800, 1.6mM for 20 minutes at room temperature.(A) Hoechst (blue) nuclear stain. (B) Phalloidin-iFluor 647 (red) F-Actin stain. (C) Ataxin 1 Antibody (D) Composite.
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