Immunoprecipitated Ataxin-1[30Q] from stably transfected CHO cells by the ataxin-1 antibody 11NQ (Skinner et al., 1997). Immunostaining of ataxin-1 was assessed with either M5 (anti-FLAG, 1:200, Sigma) or 11750 (anti-ataxin-1, 1:200). Western blot analysis was performed and probed with anti-ataxin-1 11750.
Analysis of ataxin-1 phosphorylation was performed with ataxin-1 antibody 11750. Western blot analysis was performed using the anti-polyglutamine antibody 1C2 (Chemicon) or anti-ataxin-1 11750.
The Rabbit polyclonal ataxin-1 S776 phospho-specific antibody (PN1168) was generated and affinity purified by New England Peptide.
Immunofluorescent staining of mouse cerebellum was performed using primary antibody 11750 (1:2000), 11NQ (1:2000), PN1168 (1:500), or calbindin (1:1000). The monoclonal ubiquitin antibody (R&D Systems) was used at a dilution of 1:1000. Secondary antibodies Anti-mouse- or anti-rabbit-conjugated Alexa Fluor 488 (Molecular Probes) was used at a dilution of 1:500, and anti-mouse- or anti-rabbit-conjugated Cy3 (Jackson Immunoresearch) was used at a dilution of 1:500.
The Mouse Genetics Laboratory (Univ. of MN) derived potential ataxin-1[82Q]-A776 transgenic founder animals. Homozygous spinocerebellar ataxia type 1 (SCA1) transgenic animals were generated.