Calprotectin human elisa

A licensed phlebotomist obtained blood samples (35ml) from participants between 4pm and 6:30pm. Cortisol levels in serum were determined with enzyme-linked immunosorbent assay (ELISA, Diagnostic Systems Laboratories, Webster, Texas), inter-assay coefficient of variation (CV) = 6%; intra-assay CV = 5%. Serum s100A8/A9 levels were obtained by an ultra-sensitive ELISA (Calprotectin, Human ELISA; Hycult Biotech Inc, Wayne, PA), inter-assay CV = 4.3%; intra-assay CV = 5.3%. Self-reported cancer-related distress over the past seven days was assessed using the 22-item Impact of Event Scale—Revised (IES-R) (21 ), which includes three subscales Intrusions (IES-I), Hyperarousal (IES-H), and Avoidance (IES-A). The IES-R has been used in previous studies of stress and stress management in BCa patients (20 (link), 22 ). Cronbach alpha indicated high reliability (IES-I α=.87; IES-H α=.83; IES-A α=.81).

Serum concentrations of the pro-inflammatory cytokines, interleukin 1-beta (IL-1β), Tumor Necrosis Factor-alpha (TNF-α), and IL-6 were measured using Quantikine High Sensitivity Enzyme-linked Immunosorbent Assay (ELISA) kits from R&D Systems (USA), according to manufacturer’s instructions. The assays have good reliability (IL-1β: inter-assay coefficient of reliability (CV) = 8.1% and intra-assay CV = 3.6%; IL-6: inter-assay CV = 6.5% and intra-assay CV = 4.1%; TNF-α: inter-assay CV = 6.5% and intra-assay CV =2.0%). We also quantified levels of s100A8/A9 at T0 and T2, which we have previously analyzed (Taub et al., 2019 (link)) with ultra-sensitive ELISA (HycultBiotech Calprotectin Human ELISA). The assay was conducted in accordance with the manualized protocol (inter-assay CV = 4.3%; intra-assay CV = 5.3%).

Blood samples of approximately 35ml were obtained by a licensed phlebotomist between 4pm and 6:30pm at baseline and 12-month follow-up. Same-day centrifugation was used to separate the serum from blood samples. An ultra-sensitive enzyme-linked immunosorbent assay (HycultBiotech Calprotectin Human ELISA) was conducted by trained lab personnel on the serum samples to quantify the concentration of S100A8/A9 ligands.