Migration:Article Title: Host-Specific Interplay between Foot-and-Mouth Disease Virus 3D Polymerase and the Type-I Interferon Pathway.
Article Snippet: .. For this purpose, 30 μL of these samples and 4 μL of ladder Precision Plus Protein Dual Color Standards (Bio-Rad, Hercules, CA, USA) were deposited on 10-well Bolt Gradient 4–12% Bis-Tris Plus Polyacrylamide Gel (Life Technologies) and separation of the proteins was carried out in Bolt Running Buffer (Life Technologies) diluted 1:20 in distilled water, 10 min at 120 V then 40 min at 180 V. Once migration was complete, the proteins were transferred to a nitrocellulose membrane (Life Technologies) by liquid transfer in Bolt Transfer Buffer (Life Technologies) diluted 1:20 in distilled water with 20% ethanol (Euromedex) and 0.1% antioxidant (Life), for 1 h at 30 V. The resulting nitrocellulose membranes were saturated by incubation for 30 min in 5% milk solution (Regilait, Saint-Martin-Belle-Roche; France) diluted in PBS-Tween previously prepared by mixing 7.2 L of distilled water, 800 mL PBS 10× and 8 mL of Tween20 (Euromedex). .. Half of the membranes were then subjected to an anti-FLAG antibody produced in mouse and conjugated with HRP (Sigma-Aldrich, Saint-Louis, MO, USA), used at 1:10,000, while the other half were subjected to an anti-GST antibody produced in rabbit, used at 1:2500.
Membrane:Article Title: Host-Specific Interplay between Foot-and-Mouth Disease Virus 3D Polymerase and the Type-I Interferon Pathway.
Article Snippet: .. For this purpose, 30 μL of these samples and 4 μL of ladder Precision Plus Protein Dual Color Standards (Bio-Rad, Hercules, CA, USA) were deposited on 10-well Bolt Gradient 4–12% Bis-Tris Plus Polyacrylamide Gel (Life Technologies) and separation of the proteins was carried out in Bolt Running Buffer (Life Technologies) diluted 1:20 in distilled water, 10 min at 120 V then 40 min at 180 V. Once migration was complete, the proteins were transferred to a nitrocellulose membrane (Life Technologies) by liquid transfer in Bolt Transfer Buffer (Life Technologies) diluted 1:20 in distilled water with 20% ethanol (Euromedex) and 0.1% antioxidant (Life), for 1 h at 30 V. The resulting nitrocellulose membranes were saturated by incubation for 30 min in 5% milk solution (Regilait, Saint-Martin-Belle-Roche; France) diluted in PBS-Tween previously prepared by mixing 7.2 L of distilled water, 800 mL PBS 10× and 8 mL of Tween20 (Euromedex). .. Half of the membranes were then subjected to an anti-FLAG antibody produced in mouse and conjugated with HRP (Sigma-Aldrich, Saint-Louis, MO, USA), used at 1:10,000, while the other half were subjected to an anti-GST antibody produced in rabbit, used at 1:2500.
Incubation:Article Title: Host-Specific Interplay between Foot-and-Mouth Disease Virus 3D Polymerase and the Type-I Interferon Pathway.
Article Snippet: .. For this purpose, 30 μL of these samples and 4 μL of ladder Precision Plus Protein Dual Color Standards (Bio-Rad, Hercules, CA, USA) were deposited on 10-well Bolt Gradient 4–12% Bis-Tris Plus Polyacrylamide Gel (Life Technologies) and separation of the proteins was carried out in Bolt Running Buffer (Life Technologies) diluted 1:20 in distilled water, 10 min at 120 V then 40 min at 180 V. Once migration was complete, the proteins were transferred to a nitrocellulose membrane (Life Technologies) by liquid transfer in Bolt Transfer Buffer (Life Technologies) diluted 1:20 in distilled water with 20% ethanol (Euromedex) and 0.1% antioxidant (Life), for 1 h at 30 V. The resulting nitrocellulose membranes were saturated by incubation for 30 min in 5% milk solution (Regilait, Saint-Martin-Belle-Roche; France) diluted in PBS-Tween previously prepared by mixing 7.2 L of distilled water, 800 mL PBS 10× and 8 mL of Tween20 (Euromedex). .. Half of the membranes were then subjected to an anti-FLAG antibody produced in mouse and conjugated with HRP (Sigma-Aldrich, Saint-Louis, MO, USA), used at 1:10,000, while the other half were subjected to an anti-GST antibody produced in rabbit, used at 1:2500.
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