Immunoblotting Procedures for Neuronal Proteins

SDS gel electrophoresis and Western blotting were performed as described in our previous publications (53 (link), 54 (link)). Primary antibodies against AADC (rabbit polyclonal; Synaptic Systems; 1:1000), TH mouse monoclonal, clone 2 (mouse monoclonal, clone TH-2; Sigma; 1:5000), synaptophysin (mouse monoclonal, clone 2; BD Transduction Laboratories; 1:25,000), synapsin IIa (mouse monoclonal, clone 1; BD Transduction Laboratories; 1:5000), VAMP2 (mouse monoclonal, clone 69.1; Synaptic Systems; 1:1000), VMAT-2 (rabbit polyclonal; Sigma; 1:1000), α-synuclein (mouse monoclonal, clone 42; BD Transduction Laboratories; 1:500), β-synuclein (mouse monoclonal, clone 8; BD Transduction Laboratories; 1:5000), and γ-synuclein (rabbit polyclonal; SK23 (50 (link)); 1:1000), secondary horseradish peroxidase–conjugated antimouse or anti-rabbit antibodies (GE Healthcare; 1:3000), and ECL+ system (GE Healthcare) were used for detection of target proteins.

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Ninkina N., Millership S.J., Peters O.M., Connor-Robson N., Chaprov K., Kopylov A.T., Montoya A., Kramer H., Withers D.J, & Buchman V.L. (2021). β-synuclein potentiates synaptic vesicle dopamine uptake and rescues dopaminergic neurons from MPTP-induced death in the absence of other synucleins. The Journal of Biological Chemistry, 297(6), 101375.

Publication 2021

VAMP2 α-synuclein ECL+ system

Aadc Anti antibodies Antibodies Clone Electrophoresis Horseradish peroxidase Mouse Proteins target Rabbit Synapsin Synaptophysin Synuclein Vamp2 Vmat α synuclein

Corresponding Organization : Institute of Physiologically Active Compounds

Other organizations : MRC London Institute of Medical Sciences, Imperial College London, Institute of Biomedical Chemistry

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Variable analysis

independent variables

Primary antibodies against AADC (rabbit polyclonal; Synaptic Systems; 1:1000)
TH mouse monoclonal, clone 2 (mouse monoclonal, clone TH-2; Sigma; 1:5000)
Synaptophysin (mouse monoclonal, clone 2; BD Transduction Laboratories; 1:25,000)
Synapsin IIa (mouse monoclonal, clone 1; BD Transduction Laboratories; 1:5000)
VAMP2 (mouse monoclonal, clone 69.1; Synaptic Systems; 1:1000)
VMAT-2 (rabbit polyclonal; Sigma; 1:1000)
α-synuclein (mouse monoclonal, clone 42; BD Transduction Laboratories; 1:500)
β-synuclein (mouse monoclonal, clone 8; BD Transduction Laboratories; 1:5000)
γ-synuclein (rabbit polyclonal; SK23 (50 (link)); 1:1000)

dependent variables

Target protein expression levels

control variables

SDS gel electrophoresis and Western blotting procedures as described in previous publications (53 (link), 54 (link))
Secondary horseradish peroxidase–conjugated antimouse or anti-rabbit antibodies (GE Healthcare; 1:3000)
ECL+ system (GE Healthcare) for detection of target proteins

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