This is a knockout-validated antibody summary, based on publications "Sensitivity and specificity of phospho-Ser129 α-synuclein monoclonal antibodies", as cited below [1], and "Heterogeneity in α-synuclein fibril activity correlates to disease phenotypes in Lewy body dementia" for immunohistochemistry knockout validation (figure 2a) [2]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Rabbit monoclonal IgG

Company: Abcam

Antibody: Alpha-synuclein

Catalog number: ab51253

Summary: Rabbit monoclonal IgG against a synthetic peptide within human Alpha-synuclein (aa 100 to the C-terminus). This antibody only detects alpha synuclein phosphorylated on Ser129. Reacts with rat, mouse and human. Suitable for western blot, dot blot, immunohistochemistry (paraffin), immunocytochemistry/immunofluorescence, and ELISA. Unsuitable for immunoprecipitation, flow cytometry or immunohistochemistry (frozen).

Western blot

Brain cortical tissue from wild-type and α-syn knockout mice.

5% milk in TBST for 1 hour at RT.

1:2,500 concentration in 5% milk in TBST overnight at 4°C.

1:10,000 concentration in 5% milk in TBST overnight at 4°C.

Crescendo for 1 minute, and membranes were digitally imaged on a ChemiDoc system.

Cortical tissue was dissected from wild-type and α-syn knockout mice (C57bl/6J), as indicated, and sequentially processed into buffers of increasing stringency, starting with (lane) 1) saline, 2) high-salt supplemented saline, 3) triton-X 100, and 4) SDS. Molecular weight markers are shown in left margins. The Syn1 monoclonal antibody was used to label total α-syn protein as a positive control, with a secondary-only membrane incubated with donkey anti-mouse antibody as a negative control to reveal globulins. Please see Figure 7 in the article [1].