PGEdos levels in the sera were assessed using a specific ELISA (Cayman Pharmaceuticals, Ann Arbor, MI) for the PGE2 metabolite (PGEM; 13,14-dihydro 15-keto prostaglandin A2), according to the manufacturer’s recommended protocol. Results are expressed as pg PGEM / ml have a peek at the hyperlink serum. Serum MUC1 levels were determined using the CA15-3 ELISA (Genway Biotech Inc. San Diego, CA (35)). Detection of antibody to MUC1 was carried out by a specific ELISA using BC2 (IgG) antibody that recognizes the extracellular MUC1 TR as the standard. The plate was coated with the 24-mer TAPPAHGVTSAPDTRPAPGSTAPP peptide as the capture antigen (6, 36).
Using a published HPLC assay to own IDO enzymatic activity measurement (37) while the a starting point, you will find enhanced and you will verified a sensitive HPLC assay with Ultraviolet and you will fluorescence identification that enables productive chromatographic breakup of tryptophan and you may the metabolite, kynurenine, inside the gel and cell lysate. Continue reading