Quantification of Viral Capsid Proteins

NuPAGE gels (Novex, Invitrogen, Carlsbad, CA, USA) at appropriate concentrations were used for proteins separation as described [40 (link),41 (link)]. Filters were probed with the following antibodies at reported dilution: mouse polyclonal anti-GLV-capsid protein_N-terminal (GLV-CP_NT) 1:2000; mouse polyclonal anti-GLV-capsid protein_C-terminal (GLV-CP_CT) 1:2000; mouse polyclonal anti-J17/10_B-ORF1 N-terminal (J_B-ORF1_NT) 1:2000; mouse anti-αTubulin (clone B-5-1-2, Sigma-Aldrich, Merck Life Science S.r.l., Milan, Italy), 1:10,000; rabbit polyclonal N14 (anti-g14-3-3) [40 (link)] 1:5000. Interaction was revealed by incubation with HRP-conjugated secondary Ab (Bio-Rad, Hercules, CA USA) at 1:2000–1:3000 dilution, followed by chemiluminescence (Millipore, Merck Life Science S.r.l., Milan, Italy).

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Marucci G., Zullino I., Bertuccini L., Camerini S., Cecchetti S., Pietrantoni A., Casella M., Vatta P., Greenwood A.D., Fiorillo A, & Lalle M. (2021). Re-Discovery of Giardiavirus: Genomic and Functional Analysis of Viruses from Giardia duodenalis Isolates. Biomedicines, 9(6), 654.

Publication 2021

NuPAGE gels HRP-conjugated secondary Ab

Antibodies Capsid Chemiluminescence Clone Dilution Gels Mouse Protein c Protein n Proteins Rabbit

Corresponding Organization : Istituto Superiore di Sanità

Other organizations : Leibniz Institute for Zoo and Wildlife Research, Freie Universität Berlin, Sapienza University of Rome

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Variable analysis

independent variables

NuPAGE gels (Novex, Invitrogen, Carlsbad, CA, USA) at appropriate concentrations

dependent variables

Protein separation

control variables

Mouse anti-αTubulin (clone B-5-1-2, Sigma-Aldrich, Merck Life Science S.r.l., Milan, Italy), 1:10,000
Rabbit polyclonal N14 (anti-g14-3-3) [40 (link)] 1:5000

controls

Mouse polyclonal anti-GLV-capsid protein_N-terminal (GLV-CP_NT) 1:2000
Mouse polyclonal anti-GLV-capsid protein_C-terminal (GLV-CP_CT) 1:2000
Mouse polyclonal anti-J17/10_B-ORF1 N-terminal (J_B-ORF1_NT) 1:2000

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