Raybio human egf elisa kit

Immune factors in the milk samples were determined at the Complutense University of Madrid (Spain) following the procedure described by Ruiz et al. [1 (link)]. The concentrations of innate immune factors (IL1β, IL6, IL12, IFNγ, TNFα), acquired immunity factors (IL2, IL4, IL10, IL13, IL17), chemokines (IL8, Groα, MCP1, MIP1β), and growth factors (IL5, IL7, GCSF, GMCSF, TGFβ2) were determined by magnetic beads-based multiplex immunoassays, using a Bio-Plex 200 instrument (Bio-Rad, Hercules, CA) and the Bio-Plex Pro Human Cytokine, Chemokine, and Growth Factor Assays and the Bio-Plex Pro TGF-β 3-plex Immunoassay kits (Bio-Rad). Epidermal growth factor (EGF) was determined by ELISA using the RayBio Human EGF ELISA kit (RayBiotech, Norcross, GA). The concentrations of immunoglobulins (IgA, total IgG, and IgM) in the milk samples were determined using the Bio-Plex Pro Human Isotyping Assay kit (Bio-Rad) in the Bio-Plex system instrument.
Prior to their analysis, the samples (1 mL) were processed and aliquoted as described previously [28 (link)]. Every assay was run in duplicate according to the manufacturer's instructions and standard curves were performed for each analyte.

Concentrations of Igs (IgA, IgG, and IgM), transforming growth factor-beta 2 (TGF-β₂) and hormones (adiponectin, ghrelin, and leptin) were determined in duplicate using a Bioplex 200 system instrument (Bio-Rad, Hercules, CA, USA) and the Bio-Plex Pro Human Isotyping Assay, Bio-Plex Pro Human TGF-β Assay and Bio-Plex Pro Human Diabetes Assays kits (Bio-Rad). The concentration of EGF was measured using the RayBio® Human EGF ELISA kit (RayBiotech, Norcross, GA). Every assay was performed according to manufacturer's instructions. Standard curves were performed for each analyte. The analytes were assayed in, at least, three batches for each HTST treatment.
The inter-assay coefficients of variation were below manufacturers' instructions for all the immune markers, and the lower limit of quantification (LLOQ) for every analyte in human milk were: 0.21 μg/L for IgA, 0.78 μg/L for IgM, 2.19 μg/L for IgG, 1.57 ng/L for TGF-β₂, 0.03 ng/L for EGF, 23.10 ng/L for adiponectin, 7.40 ng/L for ghrelin, and 11.45 ng/L for leptin.

Angiogenesis arrays were used to analyze differential expression of twenty human cytokines by HUVECs in response to a Bb infection. HUVECs were infected for 24 h with various Bb strains (MOI = 500). Culture supernatants were subsequently collected and analyzed by a Human Angiogenesis Antibody Array C1 as instructed by the manufacturer (RayBiotech; Peachtree Corners, GA). Results of the array were interpreted by the Human Angiogenesis Analysis Tool (RayBiotech). A RayBio Human EGF ELISA kit was subsequently used to assay changes in EGF concentrations in the HUVEC culture medium in response to Bb lysates, formaldehyde-fixed Bb or Bb-infection at 24 h post-infection or post-treatment, following the manufacturer’s instructions (RayBiotech).