Western Blot with HeLa cell extracts [25 μg/lane] from UBXD7 WT expressing or UBXD7 depleted or over-expressing HeLa cells. Total cell extracts were immunoblotted with purified anti-UBXD7 antibodies (5% milk in TBS-Tween, o/n). Binding of the primary antibody was detected using peroxidase conjugated anti-sheep IgG antibody (1:3000 dilution) followed by enhanced chemiluminescence (ECL, Amersham).
Mouse brain lysate (9.4 mg protein lysate) was immunoprecipitated with 80 μl anti- UBXD7 crosslinked to Protein A (PrA) agarose beads (dry beads). For the antibody crosslinking: 250 μg anti-UBXD7 antibody were crosslinked to 1 ml Protein A agarose beads [dry beads]. WB: load 50 μg for Input, 8% of the IP