1. Run ~50 ug of total protein on a 4-15% SDS polyacrylamide gel.
2. Transfer protein to a nitrocellulose membrane.
3. Block membrane with 1XPBS/5% non-fat milk/0.1% Tween-20 for 1 hour at room temperature (~23-27C).
4. Incubate membrane with 1:1000 dilution of NB400-114, diluted in 1XPBS/1% BSA, for 1 hour at room temperature.
5. Wash membrane once for 15 minutes, then four times for 5 minutes each, with PBST.
6. Incubate membrane with anti-rabbit IgG-HRP, diluted in 1XPBS/1% BSA, for 30 minutes-1 hour at room temperature.
7. Wash membrane once for 15 minutes, then four times for 5 minutes each, with PBST.
8. Detect cross-reacting proteins using a Chemiluminescence Reagent kit: expose ~1 minute.
NOTE: mouse liver lysates were used as a positive control for this antibody.
