Western Blot Protocol for HIP1 Antibody (NB300-204)

Western Blot Procedure

1. Run 25 ug/lane of protein (ie: HeLa whole cell extract) on a 10% SDS-PAGE gel.
2. Transfer the proteins to nitrocellulose.
3. Block the membrane with a standard blocking buffer (5% NFDM in TBST), overnight at 4 deg C.
4. Dilute the anti-HIP1 in 1% NFDM in TBST and incubate for 1hour at RT.
5. Wash the membrane, 3x 5-10 minutes.
6. Dilute the secondary antibody [anti-mouse IgG conjugated to HRP] in 1% NFDM in TBST and incubate for 1hour at RT.
7. Wash the membrane, 3x 5-10 minutes.
8. Detect the protein-antibody complex via ECL.