The entire sequence encoding UBXN-6 was amplified by PCR with a cDNA clone yk1159e09 as a template, and primers ubxn6- Cold-5 and ubxn6-Cold-3, and cloned onto pCold I, yielding pMUR7. Escherichia coli BL21(DE3) was transformed with pMUR7. His-tagged UBXN-6 was overexpressed by temperature shift-down and the addition of 1 mM isopropyl-1-thio-b-D-galactopyranoside and purified by using HisTrap HP and HiTrap Q columns (GE Healthcare). Purified His-tagged UBXN-6 was pooled and used for development of antibody. Rabbit anti-UBXN-6 antisera was devel- oped by IWAKI&CO., LTD. and purified using antigen-conjugated affinity resin.
ubxn-6