Journal: Molecular Neurodegeneration
Article Title: Autoimmune antibody decline in Parkinson’s disease and Multiple System Atrophy; a step towards immunotherapeutic strategies
Figure Lengend Snippet: Cross-binding competition ELISA assays. a , b , and c : The two site inhibition curves show distinct high and low binding components in plasma from Parkinson’s Disease ( PD-blue, squares ), Multiple System Atrophy ( MSA-green, triangles ) patients and normal control ( NC-black, circles ) subjects. Ten plasma samples from each group were pooled and incubated in 1:400 dilution with increasing concentration of α-, β-, and γ-synuclein monomers in combinations as indicated, with subsequent measurement of free NAbs by ELISA on plates coated with 10 μg/ml of α-synuclein. d - i : Percentage of inhibition (PI) of individual plasma samples with free d , e : α−/β-synuclein monomers, F , G : α−/γ-synuclein monomers; h , i : β−/γ-synuclein monomer on plates coated with 10 μg/mL of α-synuclein monomer. Plasma samples from Parkinson’s Disease (PD), Multiple System Atrophy (MSA) patients and normal controls (NC) were tested at 1:400 dilution in the presence of ( d , f , h ) 100 nM or ( e , g , i ) 10 nM α-, β-, and γ-synuclein monomers in combinations. Horizontal bars represent the mean values +/− SEM. Significance was tested using Man–Whitney’s U test ( P
Article Snippet: 96-well polystyrene microtiter plates (Nunc MaxiSorp® flat-bottom 96 well plate) were coated with either 10 μg/mL recombinant β-synuclein monomer (rPeptide,GA,USA, #S-1003-2) or 10 μg/mL recombinant γ-synuclein monomer (rPeptide,GA,USA, #S-1007-1) in ice-cold 0.1 M carbonate buffer (pH 8.5) overnight ( > 12 h) at 4 °C.
Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay, Inhibition, Incubation, Concentration Assay