Western Blot
1. Resolve proteins on a 10% SDS-PAGE.
2. Transfer to nitrocellulose.
3. Block in 5% NFDM overnight at 4C.
4. Dilute anti-GAPDH (NB 300-328) 1:5,000 in blocking buffer and incubate for 2 hours at RT.
5. Dilute secondary goat anti-mouse IgG (whole molecule) conjugated to HRP (as per manufacturer's guidelines) and incubate for 1 hour at RT.
6. Develop membrane using ECL reagent.