Purification and Characterization of ASPH Antibodies

Pharmaceutical grade ethanol was used in the in vivo experiments. The A85G6 and A85E6 monoclonal antibodies to ASPH and Humbug were generated into human recombinant proteins and purified over Protein G columns (Healthcare, Piscataway, NJ, USA) [34 (link)]. All other primary antibodies used for the duplex ELISAs were purchased from Abcam (Cambridge, MA, USA). RPLPO antibody was sourced from the Proteintech Group Inc (Chicago, IL, USA). ELISA MaxiSorp 96-well plates were purchased from Nunc (Rochester, NY, USA). Horseradish peroxidase (HRP)-conjugated secondary antibody, and Amplex Red soluble fluorophore were purchased from Invitrogen (Carlsbad, CA, USA). The SpectraMax M5 microplate reader was purchased from Molecular Devices Corp. (Sunnyvale, CA, USA). BCA reagents were obtained from Pierce Chemical Corp. (Rockford, IL, USA). All other fine chemicals were purchased from CalBiochem (Carlsbad, CA, USA), Pierce (Rockford, IL, USA), or Sigma (St. Louis, MO, USA).