ABCG8 Antibody:
Western Blot Protocol
1. Load ~50 micrograms of lysate (ie: liver membrane) onto an 8% SDS-PAGE, reducing.
2. Block the protein transferred membrane for 30 minutes in blocking buffer [5% NFDM, 10 mM Tris (pH 7.6) saline, 0.2% Tween-20] at room temperature.
3. Incubate the membrane in diluted primary anti-ABCG8 [cat# NB 400-117], 1:250 for 1 hour at room temperature.
4. Wash the membrane 3x 5 minutes in blocking buffer without milk.
5. Incubate the membrane in diluted secondary anti-Rabbit IgG conjugated to HRP.
6. Wash the membrane 3x 5 minutes in blocking buffer without milk.
7. Wash the membrane for 5 minutes in TBS.
8. Develop using West-Pico ECL reagent (Pierce).
