For Western blot, samples were transferred to PVDF membrane after SDS-PAGE using a wet transfer system.
Using standard protocols for SDS-PAGE and then Western blots, all samples were transferred to Immobilon membranes and probed with appropriate antibodies to detect the proteins of interest.
For western blot analysis, samples were transferred to PVDF membrane (Immobilon-FL, IPFL00010), blocked with Odyssey Blocking Buffer (Licor, 927 40,010), and probed with anti-Rtls antibody at 1 2500 followed by the secondary antibody anti-rabbit IRDye 800CW (Licor, 926 322111).
For Western blotting, the protein samples were transferred onto a HYBOND™-C EXTRA nitrocellulose membrane using a Trans-blot Semi-dry Transfer Cell (BioRad®).
The protein samples were transferred by western blotting onto PVDF membrane, which were blocked with PBS containing 5%(w/v) skimed milk powder (Marvel™) as described previously (McDonald et al. 2004).
The samples were transferred onto Immuno-blot PVDF membrane (Bio-Rad, Hercules) in transfer buffer (192 mM glycine; 25 mM Tris-HCl) overnight at 30V.
After separation in 12.5% SDS-PAGE, protein samples were transferred onto polyvinylidene fluoride (PVDF) membrane using blotting apparatus.
Separate gels were stained with Coomassie Blue (Pierce, Rockford, IL), or the samples were transferred to nitrocellulose using an X-Cell II Blot Module Semi-Dry Transfer unit (Invitrogen) for Western blotting.
Samples were transferred to nitrocellulose membranes and subjected to Western blot analysis.
After electrophoresis, samples were transferred to a nitrocellulose membrane using a Trans-Blot Turbo transfer system (Bio-Rad, Hercules, CA, USA).
