Western Blot with HeLa cell extracts [30 μg/lane] from UBXN8 WT expressing or UBXN8 depleted (UBXN8) HeLa cells. Total cell extracts were immunoblotted with purified anti-UBXN8 (R2824) antibodies. Binding of the primary antibody was detected using rabbit peroxidase conjugated anti-rabbit IgG antibody (1:3000 dilution) followed by enhanced chemiluminescence (ECL, Amersham).

Total cell extracts (1 mg) were immunoprecipitated with 60 μl anti- UBXN8 crosslinked to Protein A (PrA) agarose beads (50% slurry). For the antibody crosslinking: 170 μg anti-UBXN8 antibody (R2824 [1/2/3 bleed pool]) were crosslinked to 1 ml Protein A agarose beads [50% slurry]. Binding of the primary antibody was detected using rabbit peroxidase conjugated anti- rabbit IgG antibody (1:3000 dilution) followed by enhanced chemiluminescence.