Rabbit anti nop10

The purified telomerase samples (E1 and E2) from the CHAPS lysis protocol were resolved by bis-Tris SDS-PAGE. Proteins were transferred to a nitrocellulose membrane and blocked with 4% nonfat milk (Carnation) in TBS buffer (50 mM Tris pH 7.5, 150 mM NaCl) for 1 hour at room-temperature. The membranes were incubated with primary antibodies overnight at 4°C and washed with TBS before being probed with secondary antibodies (Invitrogen) in 4% nonfat milk in TBS for 1 hour at room-temperature. We blotted for TERT, TCAB1, dyskerin and GAR1 on the same membrane, and NHP2 and NOP10 on a different membrane. The small sizes of NHP2 and NOP10 required a shorter protein transfer time than the other proteins. The membranes were washed extensively with TBS and visualized on a LI-COR Odyssey imager. The primary antibodies used were rabbit anti-dyskerin (Santa Cruz Biotechnology), mouse anti-TCAB1 (Sigma), rabbit anti-GAR1 (Novus), rabbit anti-NHP2 (Proteintech), rabbit anti-NOP10 (Abcam), mouse anti-Strep (Abcam), mouse anti-tubulin (Abcam) and rabbit IgG (Sigma-Aldrich).