Characterization of Recombinant α-Synuclein

To characterize the property of recombinant A-syn, the thrombin-cleaved A-syn was subjected to gel filtration chromatography on 10/300 GL Superdex 200 column (GE healthcare) using the biologic Duoflow system (Bio-Rad). The monomeric A-syn was collected and confirmed by another gel filtration. The elution volume of A-syn was determined using standard molecular mass markers (thyroglobulin, 670 kDa; γ-globulin, 158 kDa; ovalbumin, 44 kDa; myoglobin, 17 kDa; and vitamin B12, 1.35 kDa; Bio-Rad) on the same column. Western blotting of A-syn was carried out as previously described [39 (link)].

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Lou X., Kim J., Hawk B.J, & Shin Y.K. (2017). α-Synuclein may cross-bridge v-SNARE and acidic phospholipids to facilitate SNARE-dependent vesicle docking. The Biochemical journal, 474(12), 2039-2049.

Publication 2017

Duoflow system thyroglobulin γ-globulin ovalbumin myoglobin vitamin B12

Biologic Gel filtration Molecular markers Myoglobin Ovalbumin Thrombin Thyroglobulin Vitamin b12 γ globulin

Corresponding Organization : Iowa State University

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Variable analysis

independent variables

Thrombin-cleaved A-syn

dependent variables

Elution volume of A-syn

control variables

Gel filtration chromatography on 10/300 GL Superdex 200 column (GE healthcare) using the biologic Duoflow system (Bio-Rad)
Standard molecular mass markers (thyroglobulin, 670 kDa; γ-globulin, 158 kDa; ovalbumin, 44 kDa; myoglobin, 17 kDa; and vitamin B12, 1.35 kDa; Bio-Rad)

controls

Western blotting of A-syn was carried out as previously described [39 (link)]

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