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Calprotectin

Manufactured by Hycult Biotech
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Calprotectin is a protein complex found in the cytoplasm of neutrophils and monocytes. It is a marker of inflammation and can be used to quantify the level of inflammation in the body.

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Lab products found in correlation

Rnalater, by Thermo Fisher Scientific (2 mentions) Ribopure blood kit, by Thermo Fisher Scientific (1 mentions) Iscript cdna synthesis kit, by Bio-Rad (1 mentions) Sybr green supermix, by Bio-Rad (1 mentions) Ll 37, by Hycult Biotech (1 mentions) Lactoferrin, by Hycult Biotech (1 mentions) Hnp1 3, by Hycult Biotech (1 mentions) Zm241385, by Merck Group (1 mentions) Recombinant human il 4, by R&D Systems (1 mentions) Poly 1 c pic, by InvivoGen (1 mentions) Gm csf, by R&D Systems (1 mentions) Calprotectin, by R&D Systems (1 mentions) Sil 6r, by R&D Systems (1 mentions)

Spelling variants of this product

Human Calprotectin ELISA kit (4 citations) Human Calprotectin (4 citations) Calprotectin Human ELISA (3 citations) S100A9/Calprotectin mouse ELISA kit (2 citations)

7 protocols using calprotectin

1

Serum Biomarkers of Inflammation in SpA

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Serum analysis included safety measurements (liver function, renal function, blood cell counts) as well as C-reactive protein (CRP) and erythrocyte sedimentation rate (ESR). Serum levels of matrix metalloproteinase-3 (MMP-3, Biotrak, Amersham Pharmacia Biotech) and calprotectin (Hycult Biotech), two potential biomarkers of inflammation in SpA [32 (link), 33 (link)], were measured by enzyme-linked immunosorbent assays (ELISAs) according to the manufacturer’s instructions.
Paramarta J.E., Turina M.C., Noordenbos T., Heijda T.F., Blijdorp I.C., Yeremenko N, & Baeten D. (2016). A proof-of-concept study with the tyrosine kinase inhibitor nilotinib in spondyloarthritis. Journal of Translational Medicine, 14, 308.
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2

Evaluating Gut and Systemic Inflammatory Responses

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Intestinal inflammatory responses will be assessed from fecal samples obtained on day 13 of each study arm. Fecal protein concentrations of calprotectin (Hycult Biotech) and myeloperoxidase (MPO; Eagle Biosciences) will be measured by ELISA to assess between-treatment differences in gut-level inflammatory responses. Systemic inflammatory responses also will be evaluated on day 0 and day 14 using total RNA from whole blood samples by qRT-PCR to assess within- and between-treatment differences in expression levels of TLR4/NFκB signaling genes (TLR4,CD14,MD2,myeloid differentiation primary response 88,p65,IL-1,IL-6,IL-8,TNFα,monocyte chemoattractant protein-1) as we have described (60 (link)). In brief, blood samples will be mixed with RNAlater (Thermo Fisher) prior to cryogenic preservation. RNA will be isolated using a RiboPure-Blood Kit (Life Technologies) and RNA purity will be assessed at 260 nm/280 nm and 260 nm/230 nm on a BioSpec-nano spectrophotometer (Shimadzu). cDNA will then be synthesized for qRT-PCR analysis using an iScript cDNA synthesis kit (Bio-Rad) for the measurement of target genes using SYBR Green Supermix (Bio-Rad).
Quarles W.R., Pokala A., Shaw E.L., Ortega-Anaya J., Hillmann L., Jimenez-Flores R, & Bruno R.S. (2020). Alleviation of Metabolic Endotoxemia by Milk Fat Globule Membrane: Rationale, Design, and Methods of a Double-Blind, Randomized, Controlled, Crossover Dietary Intervention in Adults with Metabolic Syndrome. Current Developments in Nutrition, 4(9), nzaa130.
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3

Biomarker Serum Level Analysis

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We determined the serum levels of the following candidate biomarkers by ELISA as indicated by the manufacturers: high-sensitivity (hs)-CRP (Research & Diagnostic Systems, Inc. Minneapolis), IL-6 (Research & Diagnostics Systems, Inc.), PTX-3 (Research & Diagnostic Systems, Inc.), alpha-2-MG (Abcam, Cambridge, UK), MMP-3 (Biotrak, Amersham Pharmacia Biotech, Buckinghamshire, UK), VEGF (Research & Diagnostics Systems, Inc.), and Calprotectin (Hycult Biotech, the Netherlands). All assays were performed in duplicate.
Turina M.C., Yeremenko N., Paramarta J.E., De Rycke L, & Baeten D. (2014). Calprotectin (S100A8/9) as serum biomarker for clinical response in proof-of-concept trials in axial and peripheral spondyloarthritis. Arthritis Research & Therapy, 16(4), 413.
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4

Quantification of Oral Antimicrobial Peptides

Cited 1 time
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A range of antimicrobial peptides which are commonly associated with the oral cavity were selected for assessment in our patient’s saliva [32 (link)–34 (link)]. The following AMPs were assessed by ELISA, LL-37, Calprotectin, Lactoferrin, HNP1-3 (Hycult biotech, The Netherlands), Histatin 5 (Stratech scientific, Suffolk, UK) and Beta defensin 1 (BD1) (Peprotech, London, UK) as per manufacturer’s instructions. Clarified saliva samples were diluted 1:5 in assay buffer (PBS, 0.5% BSA, 0.1% Tween20). Results were calculated using a 4-parameter curve fit, quantifying colorimetric changes at 630 nm (BMG-Labtech, Ortenberg, Germany).
O’Donnell L.E., Robertson D., Nile C.J., Cross L.J., Riggio M., Sherriff A., Bradshaw D., Lambert M., Malcolm J., Buijs M.J., Zaura E., Crielaard W., Brandt B.W, & Ramage G. (2015). The Oral Microbiome of Denture Wearers Is Influenced by Levels of Natural Dentition. PLoS ONE, 10(9), e0137717.
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5

Immunomodulatory Factors Analysis

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Recombinant human IL-4 and GM-CSF were purchased from R&D Systems (Wiesbaden, Germany). LPS from Salmonella Minnesota (smooth form) was provided by U. Seydel (Borstel, Germany), R848 (Resiquimod, imidazoquinoline compound) was from ALEXIS (Lausen, Switzerland). Poly I:C (PIC) was obtained from InvivoGen (San Diego, CA, USA). IDO inhibitor 1-methyl-tryptophan (1MT), ADO-receptor antagonist ZM-241385 and ADO from Sigma-Aldrich (Taufkirchen, Germany), Calprotectin from Hycult Biotech (Uden, Netherlands), kynurenine and LILRA3 from Novoprotein (Summit, USA), α-IL-1β (eBioscience, Frankfurt/Main, Germany).
Giesbrecht K., Eberle M.E., Wölfle S.J., Sahin D., Sähr A., Oberhardt V., Menne Z., Bode K.A., Heeg K, & Hildebrand D. (2017). IL-1β As Mediator of Resolution That Reprograms Human Peripheral Monocytes toward a Suppressive Phenotype. Frontiers in Immunology, 8, 899.
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6

Evaluating Intestinal and Systemic Inflammation

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The interactive effects of GTE and MetS status on intestinal and systemic inflammation will be evaluated at day 28. From the collected fecal samples, protein concentrations of calprotectin (Hycult Biotech) and myeloperoxidase (Eagle Biosciences) will be determined using commercially available ELISA kits as described [51 (link)]. Systemic inflammatory responses will be evaluated from plasma by determining protein concentrations of high-sensitivity C-reactive protein, IL-6 and -8, TNFα, and myeloperoxidase using separate commercially available ELISA kits (obtained from Biocheck, Inc or R&D Systems). In addition, expression levels of genes involved in TLR4/NFκB signaling (TLR4, MyD88, p65, IL-6 and -8, TNFα, MPO, MCP-1) will be assessed from RNA isolated from peripheral blood mononuclear cells using qRT-PCR as we described [18 (link)].
Hodges J.K., Zhu J., Yu Z., Vodovotz Y., Brock G., Sasaki G.Y., Dey P, & Bruno R.S. (2019). Intestinal-level anti-inflammatory bioactivities of catechin-rich green tea: Rationale, design, and methods of a double-blind, randomized, placebo-controlled crossover trial in metabolic syndrome and healthy adults. Contemporary Clinical Trials Communications, 17, 100495.
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7

Quantifying sIL-6R and Calprotectin in GCF

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Both sIL-6R and calprotectin concentrations in GCFs were measured using commercialized enzyme-linked immunosorbent assay Kit (ELISA) (sIL-6R: R&D Systems, Minneapolis, MN; calprotectin: Hycult Biotech, PB Uden, the Netherlands) according to the manufacturer's instruction.
Kajiura Y., Lew J.H., Ikuta T., Nishikawa Y., Kido J.I., Nagata T, & Naruishi K. (2017). Clinical significance of GCF sIL-6R and calprotectin to evaluate the periodontal inflammation. Annals of clinical biochemistry, 54(6).
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