Serum concentration of Intestinal Fatty Acids-Binding Protein (I-FABP) and the soluble IL-33 receptor sST2 were measured by ELISA (Human Intestinal Fatty Acid Binding Protein I-FABP ELISA kit; CUSABIO, Houston, TX, USA and Human ST2/IL-33R Quantikine ELISA kit, R&D Systems, Minneapolis, MN, USA), following each manufacturer protocols. Also, fecal levels of secretory IgA (sIgA), calprotectin and lactoferrin were determined by ELISA, employing the following kit: Secretory IgA ELISA, sIgA (ALPCO, Salem, NH, USA), Human Lactoferrin and Human Calprotectin (both Hycult Biotech, Uden, the Netherlands) according to manufacturer instructions.
Human calprotectin
Manufactured by Hycult Biotech
Sourced in United States, China
Calprotectin is a protein complex found in neutrophils and other immune cells. It is a sensitive marker of inflammation and can be used to measure the degree of intestinal inflammation.
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Lab products found in correlation
4 protocols using human calprotectin
1
Serum and Fecal Biomarkers for Intestinal Health
2
Quantification of Antimicrobial Peptides
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AMP levels were measured using commercially available enzyme‐linked immunosorbent assay (ELISA) kits, following the manufacturers' instructions (human beta‐defensin 124: catalog# SEQ656Hu; human ribonuclease A3: catalog# SEB758Hu; all Uscn Life Science, Wuhan, China; human LL‐37, cathelicidin: catalog# HK321; human calprotectin: catalog# HK325; all Hycult Biotech, Uden, the Netherlands; human angiogenin, RNase5: catalog# DY265 R&D Systems, Minneapolis, MN, USA; and rat calprotectin: catalog# HK321; Immundiagnostik AG, Bensheim, Germany). Optical density values were measured at 450 nm on an ELISA plate reader (Victor3 multilabel plate reader; PerkinElmer, Waltham, MA, USA).
3
Circulating Neutrophil Activation Markers
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Levels of circulating neutrophil activation markers calprotectin, myeloperoxidase (MPO), elastase, bactericidal permeability increasing protein (BPI), and LCN-2 were measured by solid-phase enzyme-linked immunosorbent assays (ELISA) based on the sandwich principle, according to the manufacturer’s instructions (Hycult Biotech, Uden, The Netherlands; Human calprotectin, Catalog #HK379; Human MPO, Catalog # HK324; Human elastase, Catalog # HK319; Human BPI, Catalog # HK314; Human LCN-2, Catalog # HK330). All plasma samples were analyzed in duplicate in the same run. The intra- and inter-assay coefficients of variance of the various assays were <10%. Clinical laboratory data including circulating C-reactive protein (CRP), albumin, neutrophils (%), and lymphocytes (%) were measured in the clinical setting. For some of the patients, these clinical data were not available, the exact number of the studied patients for these data is indicated in each figure legend.
4
Quantifying Inflammation and Digestive Biomarkers
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Plasma and stool lipocalin-2 concentrations were determined using a commercially available kit to perform a sandwich ELISA for human lipocalin-2 (Human Lipocalin-2/NGAL R&D Systems, Minneapolis, MN). Plasma samples were diluted 1:1,000 using the reagent diluent buffer provided in the kit. Fecal samples were diluted 1:10,000 by serial dilution. The first dilution of 1:50 used fecal extraction buffer (0.1 M Tris HCL [8.0], 0.15 M NaCl, 1 M urea, 10 mM CaCl2, 0.1 M citric acid monohydrate, 5 g/L BSA, and 0.25 mM Thimerosal). Subsequent dilutions used the reagent diluent buffer provided in the kit.
Fecal calprotectin concentrations were determined using a commercially available kit to perform a sandwich ELISA for Human Calprotectin (Human Calprotectin HycultBiotech, Wayne, PA). Samples were diluted 1:5,000 by serial dilution.
Fecal A1AT concentrations were determined using a commercially available kit to perform a sandwich ELISA for fecal alpha-1 antitrypsin (Alpha-1 Antitrypsin Immunochrom, Heppenheim, Germany). Samples were diluted 1:12,500 by serial dilution.
For each assay, a standard curve was done on every plate and samples were run in duplicate. A coefficient of variation was less than 15% for each duplicate.
Fecal calprotectin concentrations were determined using a commercially available kit to perform a sandwich ELISA for Human Calprotectin (Human Calprotectin HycultBiotech, Wayne, PA). Samples were diluted 1:5,000 by serial dilution.
Fecal A1AT concentrations were determined using a commercially available kit to perform a sandwich ELISA for fecal alpha-1 antitrypsin (Alpha-1 Antitrypsin Immunochrom, Heppenheim, Germany). Samples were diluted 1:12,500 by serial dilution.
For each assay, a standard curve was done on every plate and samples were run in duplicate. A coefficient of variation was less than 15% for each duplicate.
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