Figure 1. Western blot analysis of RAD50 using anti-RAD50 antibody (MBS1754714).

Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel)/90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.

Lane 1: human Jurkat whole cell lysates,
Lane 2: human MCF-7 whole cell lysates,
Lane 3: human Hela whole cell lysates,
Lane 4: human K562 whole cell lysates,
Lane 5: human A549 whole cell lysates,
Lane 6: human Caco-2 whole cell lysates,
Lane 7: human Daudi whole cell lysates,
Lane 8: human SH-SY5Y whole cell lysates.

After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-RAD50 antigen affinity purified polyclonal antibody (#MBS1754714) at 0.5 ug/mL overnight at 4 degree C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for RAD50 at approximately 154 kDa. The expected band size for RAD50 is at 154 kDa.