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Calprotectin elisa assay kit

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The Calprotectin ELISA Assay Kit is a laboratory tool used to quantitatively measure the level of calprotectin, a protein found in the body. The kit provides the necessary reagents and materials to perform enzyme-linked immunosorbent assay (ELISA) analysis of calprotectin levels in biological samples.

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Lab products found in correlation

Human kits, by R&D Systems (2 mentions) Epoch plate reader, by Agilent Technologies (2 mentions) Spectramax i3x, by Molecular Devices (2 mentions)

8 protocols using calprotectin elisa assay kit

1

Fecal Biomarker Quantification Protocol

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Stool specimens were allowed to thaw at room temperature and were diluted 7-fold in buffer with protease inhibitors (RIPA, radioimmunoprecipitation assay buffer). The samples were vortexed, centrifuged and the supernatants were used to measure the biomarkers. The fecal myeloperoxidase (MPO), lactoferrin (FL), lipocalin-2 (Lcn-2) and calprotectin (FC) concentrations were measured using commercially available kits that employed polyclonal antibody-based enzyme-linked immunosorbent assay (ELISA) methods, according the manufacturer’s instructions. MPO and Lcn-2 levels were determined using human kits from R&D systems, Inc (Minneapolis, USA). The FL concentration was analyzed using a BD (IBD-SCAN); TechLab Inc. (Blacksburg, USA). The FC concentration was tested using a Calprotectin Elisa Assay Kit, Eagle Biosciences, Inc. (Nashua, USA). Total protein of each sample was assayed using the BCA Protein Assay Kit from Pierce (Pittsburgh, PA). The absorption was measured using an Epoch plate reader, Bio-tek Instruments, Inc. (USA). Units were expressed as ng/mg of stool or as ng/µg protein for protein normalized data.
Prata M.D., Havt A., Bolick D., Pinkerton R., Lima A, & Guerrant R. (2016). Comparisons between myeloperoxidase, lactoferrin, calprotectin and lipocalin-2, as fecal biomarkers of intestinal inflammation in malnourished children. Journal of translational science, 2(2), 134-139.
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2

Measuring Fecal Calprotectin Levels

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Fecal calprotectin was measured using the Calprotectin ELISA Assay Kit (Eagle Biosciences). Duplicate portions of stool (50–100mg) were weighed and processed according to the manufacturer’s instructions. Absorbance was measured with a SpectraMax® i3x (Molecular Devices). Fecal calprotectin (μ g/g feces) was calculated using a 7-point standard curve. The assay’s reported normal cut-off is <43.2 μg/g.
Reasoner S.A., Bernard R., Waalkes A., Penewit K., Lewis J., Sokolow A.G., Brown R.F., Edwards K.M., Salipante S.J., Hadjifrangiskou M, & Nicholson M.R. (2023). Longitudinal Profiling of the Intestinal Microbiome in Children with Cystic Fibrosis Treated with Elexacaftor-Tezacaftor-Ivacaftor. medRxiv.
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3

Fecal Calprotectin in Pediatric CDI

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Fecal calprotectin levels were measured on twenty-five pediatric patients diagnosed with CDI. Subjects were enrolled at two tertiary-care children’s hospitals (Monroe Carrell Jr. Children’s Hospital, Nashville, Tennessee and Texas Children’s Hospital, Houston, Texas). The Institutional Review Boards of the participating institutions reviewed and approved the study (IRB 130315). Consent was obtained from all subjects in the study. Hospital laboratory records were used to identify individuals who tested positive for C. difficile at the time of diagnosis. Both hospitals employ molecular-based assays to test stool for the presence of toxigenic C. difficile. Children aged twelve months through eighteen years with CDI were eligible for enrollment. CDI was defined as a positive C. difficile diagnostic test and diarrhea. Subjects were excluded from the study if an alternative enteropathogen was identified by routine clinical testing requested by the primary medical team. Fecal samples were thawed, normalized to weight, and fecal calprotectin was measured using the Calprotectin ELISA Assay Kit (Eagle Biosciences, Nashua, NH).
Zackular J.P., Moore J.L., Jordan A.T., Juttukonda L.J., Noto M.J., Nicholson M.R., Crews J.D., Semler M.W., Zhang Y., Ware L.B., Washington M.K., Chazin W.J., Caprioli R.M, & Skaar E.P. (2016). Dietary Zinc Alters the Microbiota and Decreases Resistance to Clostridium difficile Infection. Nature medicine, 22(11), 1330-1334.
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4

Measuring Fecal Calprotectin via ELISA

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Human fecal calprotectin was measured using the Calprotectin ELISA Assay Kit (Eagle Biosciences cat. CAL35-K01) following manufacturer’s directions. Between 50 and 100 mg of pulverized human stool was used for each assayed sample.
Wilson N.G., Hernandez-Leyva A., Rosen A.L., Jaeger N., McDonough R.T., Santiago-Borges J., Lint M.A., Rosen T.R., Tomera C.P., Bacharier L.B., Swamidass S.J, & Kau A.L. (2023). The gut microbiota of people with asthma influences lung inflammation in gnotobiotic mice. iScience, 26(2), 105991.
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5

Inflammatory Biomarkers Evaluation in Systemic and Intestinal Conditions

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We evaluated both systemic inflammation and local intestinal inflammation by measuring the levels of five inflammatory biomarkers (i.e., interleukin-6 (IL-6), interleukin-8 (IL-8), immunoglobulin A (IgA), immunoglobulin M (IgM), immunoglobulin G (IgG)) in the blood, as well as fecal calprotectin (FCP). The concentrations of IgA, IgM, IgG, IL-6, and IL-8 were quantified in the serum sample using the Merck MILLIPLEX®® Protein Multifactor Liquid Chromatography Panel (Human Cytokine Autoantibody Panel). The assay was performed by Beijing Huatai Yikang Biotechnology Co. The quantification of FCP was performed using the Calprotectin ELISA Assay Kit (Eagle BioSciences, NH, USA) through an enzyme-linked immunosorbent assay (ELISA) method.
Wang D., Meng S., Li J., Zhao J., Wang Y., Du M., Wang Y., Lu W, & Zhu Y. (2023). Associations of Adherence to the 2018 World Cancer Research Fund and the American Institute for Cancer Research Dietary Recommendations with Gut Microbiota and Inflammation Levels. Nutrients, 15(17), 3705.
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6

Fecal Biomarker Quantification Protocol

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Stool specimens were allowed to thaw at room temperature and were diluted 7-fold in buffer with protease inhibitors (RIPA, radioimmunoprecipitation assay buffer). The samples were vortexed, centrifuged and the supernatants were used to measure the biomarkers. The fecal myeloperoxidase (MPO), lactoferrin (FL), lipocalin-2 (Lcn-2) and calprotectin (FC) concentrations were measured using commercially available kits that employed polyclonal antibody-based enzyme-linked immunosorbent assay (ELISA) methods, according the manufacturer’s instructions. MPO and Lcn-2 levels were determined using human kits from R&D systems, Inc (Minneapolis, USA). The FL concentration was analyzed using a BD (IBD-SCAN); TechLab Inc. (Blacksburg, USA). The FC concentration was tested using a Calprotectin Elisa Assay Kit, Eagle Biosciences, Inc. (Nashua, USA). Total protein of each sample was assayed using the BCA Protein Assay Kit from Pierce (Pittsburgh, PA). The absorption was measured using an Epoch plate reader, Bio-tek Instruments, Inc. (USA). Units were expressed as ng/mg of stool or as ng/µg protein for protein normalized data.
Prata M.D., Havt A., Bolick D., Pinkerton R., Lima A, & Guerrant R. (2016). Comparisons between myeloperoxidase, lactoferrin, calprotectin and lipocalin-2, as fecal biomarkers of intestinal inflammation in malnourished children. Journal of translational science, 2(2), 134-139.
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7

Fecal Calprotectin Quantification Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols
Fecal calprotectin was measured using the Calprotectin ELISA Assay Kit (Eagle Biosciences). Duplicate portions of stool (50–100 mg) were weighed and processed according to the manufacturer’s instructions. Absorbance was measured with a SpectraMax i3x (Molecular Devices). Fecal calprotectin (µg/g feces) was calculated using a seven-point standard curve. The assay’s reported normal cutoff is <43.2 µg/g.
Reasoner S.A., Bernard R., Waalkes A., Penewit K., Lewis J., Sokolow A.G., Brown R.F., Edwards K.M., Salipante S.J., Hadjifrangiskou M, & Nicholson M.R. (2024). Longitudinal profiling of the intestinal microbiome in children with cystic fibrosis treated with elexacaftor-tezacaftor-ivacaftor. mBio, 15(2), e01935-23.
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8

Fecal Calprotectin in Pediatric CDI

Check if the same lab product or an alternative is used in the 5 most similar protocols
Fecal calprotectin levels were measured on twenty-five pediatric patients diagnosed with CDI. Subjects were enrolled at two tertiary-care children’s hospitals (Monroe Carrell Jr. Children’s Hospital, Nashville, Tennessee and Texas Children’s Hospital, Houston, Texas). The Institutional Review Boards of the participating institutions reviewed and approved the study (IRB 130315). Consent was obtained from all subjects in the study. Hospital laboratory records were used to identify individuals who tested positive for C. difficile at the time of diagnosis. Both hospitals employ molecular-based assays to test stool for the presence of toxigenic C. difficile. Children aged twelve months through eighteen years with CDI were eligible for enrollment. CDI was defined as a positive C. difficile diagnostic test and diarrhea. Subjects were excluded from the study if an alternative enteropathogen was identified by routine clinical testing requested by the primary medical team. Fecal samples were thawed, normalized to weight, and fecal calprotectin was measured using the Calprotectin ELISA Assay Kit (Eagle Biosciences, Nashua, NH).
Zackular J.P., Moore J.L., Jordan A.T., Juttukonda L.J., Noto M.J., Nicholson M.R., Crews J.D., Semler M.W., Zhang Y., Ware L.B., Washington M.K., Chazin W.J., Caprioli R.M, & Skaar E.P. (2016). Dietary Zinc Alters the Microbiota and Decreases Resistance to Clostridium difficile Infection. Nature medicine, 22(11), 1330-1334.
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