Total proteins were separated on a 12% SDS/PAGE gel, transferred to nitrocellulose membrane (Whatman).
Fractions were dialyzed against PBS, loaded on a SDS-PAGE gel, transferred to nitrocellulose membranes and incubated with specific rabbit policlonal antibodies to confirm its identity.
About 20 µg/sample of total RNA were separated in a formaldehyde/agarose gel, transferred onto nylon membranes and hybridized to a non-radioactively labeled probe from the coding region of chimeric polyprotein gene (Dm-AMP1+Rs-AFP2).
Samples were then fractionated on 10%% acrylamide gel, transferred to a PVDF membrane (Bio-Rad), and incubated with specific primary antibodies followed by the corresponding peroxidase-conjugated secondary antibodies.
Samples were analyzed by SDS-polyacrylamide gel, transferred to nitrocellulose membranes and autoradiographed.
For immunoblotting, protein samples were separated on a 6% SDS-PAGE gel, transferred to Immobilon™ Transfer Membrane (Millipore, Bedford, MA).
The restricted fragments were separated by 0.8% agarose gel, transferred to nylon membranes and hybridized with 32P-labeled probes of hDHFR and maebl M2 domain.
Proteins were resolved on a 12% high-TEMED SDS polyacrylamide gel, transferred to a membrane, and analysed by western blots with anti-HA conjugated to Alexa800 fluorophore (Rockland).
Samples were loaded into a 7% polyacrylamide gel, transferred onto PVDF membrane and blocked for 1.5 hours in 5% dry milk∶TBS-Tween solution.
Then, proteins were separated using SDS-PAGE on an 8% or 15% gel, transferred onto nitrocellulose membrane by PowerPac (BIO-RAD).
Lysates were separated on a 12% polyacrylamide gel, transferred to Hybond-P membranes (Amersham Pharmacia Biotech, Buckinghamshire, England) and probed with primary antibodies.
