Equal amounts of lysates were loaded onto Mini-Protean TGX (4-20%) SDS-PAGE gels (BioRad). All blots except phospho-MTFR1L were transferred to nitroceullose membranes (BioRad) with a Trans-Blot Turbo and blocked for one hour in Intercept Buffer (Li-COR). Membranes were then incubated in their respective primary antibodies in blocking buffer at 4°C overnight. Blots probing for phospho-MTFR1L we transferred to polyvinylidene difluoride membranes (PVDF, Immobilion) and blocked in 5% fat-free dry milk in TBS-T for 1 hour before primary antibody incubation for four days at 4°C. Membranes were incubated in Li-Cor fluorescence-coupled secondary antibodies for 1 hour at room temperature prior to visualization with a Li-Cor Odyssey Blot Imager.
Nitroceullose membranes
Manufactured by Bio-Rad
Nitrocellulose membranes are porous sheets made of nitrocellulose, a material derived from cellulose. They are commonly used in various laboratory techniques, such as Western blotting, dot blotting, and immunodetection, to immobilize and detect specific proteins or other biomolecules.
Automatically generated - may contain errors
2 protocols using nitroceullose membranes
1
Western Blotting Protocol for Protein Analysis
2
Western Blot Protocol
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Equal amounts of lysates were loaded onto Mini-Protean TGX (4-20%) SDS-PAGE gels (BioRad). All blots except phospho-MTFR1L were transferred to nitroceullose membranes (BioRad) with a Trans-Blot Turbo and blocked for one hour in Intercept Buffer (Li-COR). Membranes were then incubated in their respective primary antibodies in blocking buffer at 4 °C overnight. Blots probing for phospho-MTFR1L we transferred to polyvinylidene difluoride membranes (PVDF, Immobilion) and blocked in 5% fat-free dry milk in TBS-T for 1 hour before primary antibody incubation for four days at 4 °C. Membranes were incubated in Li-Cor fluorescence-coupled secondary antibodies for 1 hour at room temperature prior to visualization with a Li-Cor Odyssey Blot Imager.
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