Nupage tris acetate sds buffer

Protein samples were resolved by SDS–polyacrylamide gel electrophoresis on NuPAGE Novex 4–12% Bis-Tris gels (NP0321 and WG1402A, ThermoFisher Scientific) with NuPAGE MOPS SDS buffer (NP0001, ThermoFisher Scientific), or NuPAGE Novex 3–8% Tris-Acetate gels (EA0375BOX and WG1602BOX, ThermoFisher Scientific) with NuPAGE Tris-Acetate SDS buffer (LA0041, ThermoFisher Scientific). Resolved proteins were either stained with colloidal Coomassie blue dye (‘Instant Blue’, Expedion), or were transferred onto a nitrocellulose iBlot membrane (Invitrogen) with the iBlot Dry Transfer System (Invitrogen).
Antibodies used for protein detection in this study are described in Appendix 1-key resources table. Conjugates to horseradish peroxidase of anti-sheep IgG from donkey (Sigma, A3415), anti-rabbit IgG from donkey (GE Healthcare, NA934), or anti-goat IgG from rabbit (Sigma, A5420) were used as secondary antibodies before the detection of chemoluminescent signals on Hyperfilm ECL (Amersham, GE Healthcare) using ECL Western Blotting Detection Reagent (GE Healthcare).

Protein samples were resolved by SDS–PAGE using the following systems: NuPAGE Novex 4 - 12% Bis-Tris gels (NP0321 and WG1402A, ThermoFisher Scientific) with NuPAGE MOPS SDS buffer (NP0001, ThermoFisher Scientific) or NuPAGE MES SDS buffer (NP0002, ThermoFisher Scientific); NuPAGE Novex 3 - 8% Tris-Acetate gels (EA0375BOX and WG1602BOX, ThermoFisher Scientific) with NuPAGE Tris-Acetate SDS buffer (LA0041, ThermoFisher Scientific). The resolved proteins were either stained with colloidal Coomassie blue dye (‘Instant Blue’, ab119211, Abcam), or else were transferred onto a nitrocellulose iBlot2 membrane (2NR290123-01, ThermoFisher Scientific) with the iBlot2 Dry Transfer System (IB21001, Invitrogen), according to the manufacturer’s instructions.
The antibodies used for immunoblotting in this study are described in Table S5. Chemoluminescent signals were detected by azure-biosystems 300Q with ECL Western Blotting Detection Reagent (17039552, GE Healthcare).

Protein samples were resolved by SDS–PAGE using the following systems: NuPAGE Novex 4 ‐ 12% Bis‐Tris gels (NP0321 and WG1402A, Thermo Fisher Scientific) with NuPAGE MOPS SDS buffer (NP0001, Thermo Fisher Scientific) or NuPAGE MES SDS buffer (NP0002, Thermo Fisher Scientific); NuPAGE Novex 3 ‐ 8% Tris‐Acetate gels (EA0375BOX and WG1602BOX, Thermo Fisher Scientific) with NuPAGE Tris‐Acetate SDS buffer (LA0041, Thermo Fisher Scientific). The resolved proteins were either stained with colloidal Coomassie blue dye (“Instant Blue”, ab119211, Abcam), or else transferred onto a nitrocellulose iBlot membrane (NRO11020‐01, Thermo Fisher Scientific) with the iBlot Dry Transfer System (IB1001, Invitrogen), according to the manufacturer’s instructions.
The antibodies used for immunoblotting in this study are described in Appendix Table S1. Chemiluminescent signals were detected on Hyperfilm ECL (Amersham, 66601, GE Healthcare) using ECL Western Blotting Detection Reagent (17039552, GE Healthcare). New antibodies that were generated in this study are validated in Appendix Fig S4.