Rabbit anti adpr

For WB analysis, proteins were separated via SDS-page on a 12% SDS-polyacrylamide gel at 120V. A wet-transfer into a PVDF membrane was performed at 30 V over-night and membranes were blocked with 5% milk in TBS-T for 1 h at RT. Primary antibodies were diluted in 1% milk in TBST and incubated at 4°C over-night. After 3 washes, the secondary antibody, diluted in TBST, was incubated for 1 h at RT. After another 3 washes, specific proteins/bands were visualized with the Odyssey infrared imaging system (LI-COR). The following primary and secondary antibodies were used for WB analysis at the indicated concentrations: rabbit anti-HA (Abcam, 1:1000), rabbit anti-GST Z5 (Santa-Cruz, 1:1000), rabbit anti-CoxIV (Abcam, 1:1000), mouse anti-ATP5a (Abcam, 1:1000), rabbit anti-ADPR (N/A, 1:500), rabbit anti-ADPR (CST, 1:5000), IRDye 800CW goat anti-rabbit IgG (1:15,000, LI-COR, P/N 925-32211), and IRDye 680RD Goat anti-Mouse IgG (1:15,000, LI-COR, P/N 925-68070). For dot blot analysis, auto-modified proteins or isolated PAR chains were vacuum-blotted onto a nitrocellulose membrane, that was further blocked in milk and stained with antibodies as described above.