Protocol specific for RNase L Antibody (NB100-351)

Western Blot

1. Proteins are separated on a 10% SDS-PAGE gel.

2. Proteins are transferred to Immobilon-P membranes (Millipore Co.).

3. Following the protein transfer, the membrane is blocked with PBS-T [PBS + 0.07% Tween-20]+ 5% NFDM.

4. Anti-RNase L [cat# NB 100-351] is diluted 1:1,000, in blocking buffer and incubated for 2 hour at room temperature, gently shaking.

5. The membrane is then washed, 3 times with PBS-T, 5 minutes each.

6. Secondary antibody is incubated for 1 hour at room temperature, gently shaking.

7. The membrane is then washed, 4 times with PBS-T, 5 minutes each.

8. Membrane is developed using ECL reagents.

NOTE: Cell extracts of insect cells expressing human RNase L (0.1 ug) and Hey 1B (100 ug) were used as positive controls for this antibody.