Western blot analysis of HELLS was performed by loading whole cell lysate in 1X LDS sample buffer with 2-ME from 5 x 10^5 mouse transformed pre-B cells. Protein Ladder was Spectra™ Multicolor High Range Protein Ladder (Product # 26625). Samples were loaded onto a 3-8% Tris-acetate polyacrylamide gel (Product # EA03785BOX). Proteins were transferred to nitrocellulose membrane with wet/tank transfer. Membrane was blocked in 5% milk/TBST. The membrane was probe with a polyclonal anti-HELLS antibody (Product # PA5-76833) at a dilution of 1:1000 in 2% milk/TBST at 4°C overnight, followed by a secondary antibody HRP-anti-rabbit at a dilution of 1:5000 at room temperature for 1 hour. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # PI-32209).