Western Blot Protocol for ATM Antibody (NB100-306)

Western Blot Procedure

1) Resolve protein samples on a 6% SDS-PAGE gel at 185V for ~1.5 hours.

2) Transfer to PVDF membranes at 25V for ~1.5 hours.

3) Block the membrane with TBST+BSA and goat serum for 1 hour at RT.

4) Dilute primary antibody (NB 100-306) to 1:1,000 in blocking buffer.

5) Incubate membrane overnight at 4 degrees Celcius in diluted anti-ATM-kinase.

6) Wash 3 times ten minutes on a shaker.

7) Incubate membranes with HRP conjugated anti-mouse IgG for 1 hour (RT), diluted in blocking buffer.

8) Wash 3 times ten minutes on a shaker.

9) Add ECL reagent, as per kit directions, and expose for 1-5 seconds.