Human Zonulin ELISA Kit (DEIA2225)

Regulatory status: For research use only, not for use in diagnostic procedures.

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Size
96T
Sample
serum, plasma, saliva, urine, serum-free cell culture supernates
Species Reactivity
Human
Intended Use
For the quantitative determination of human Haptoglobin concentrations in serum-free cell culture supernates, serum, plasma, saliva, and urine.
Contents of Kit
1. Human Haptoglobin Microplate: 96 well polystyrene microplate (12 strips of 8 wells)
2. Human Haptoglobin Conjugate: 21 mL
3. Human Haptoglobin Standard: 400 ng
4. Assay Diluent CD2-210: 2 vials (11 mL/vial)
5. Calibrator Diluent CD6-78: 21 mL
6. Wash Buffer Concentrate: 21 mL
7. Color Reagent A: 12 mL
8. Color Reagent B: 12 mL
9. Stop Solution: 6 mL
10. Plate Sealers: 4 adhesive strips
Storage
Store the unopened kit at 2-8°C. Do not use past kit expiration date.
Precision
Intra-assay Precision (Precision within an assay)
Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.

Inter-assay Precision (Precision between assays)
Three samples of known concentration were tested in twenty separate assays to assess inter-assay precision. Assays were performed by at least three technicians using two lots of components.
Sensitivity
Twenty-five assays were evaluated and the minimum detectable dose (MDD) of human Haptoglobin ranged from 0.031-0.529 ng/mL. The mean MDD was 0.192 ng/mL.
The MDD was determined by adding two standard deviations to the mean optical density value of twenty zero standard replicates and calculating the corresponding concentration.
Standard Curve

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References


Effects of PrObiotics on the Symptoms and Surgical ouTComes after Anterior REsection of Colon Cancer (POSTCARE): A Randomized, Double-Blind, Placebo-Controlled Trial

JOURNAL OF CLINICAL MEDICINE

Authors: Park, In Ja; Lee, Ju-Hoon; Kye, Bong-Hyeon; Oh, Heung-Kwon; Cho, Yong Beom; Kim, You-Tae; Kim, Joo Yun; Sung, Na Young; Kang, Sung-Bum; Seo, Jeong-Meen; Sim, Jae-Hun; Lee, Jung-Lyoul; Lee, In Kyu

We investigated microbiota changes following surgical colon cancer resection and evaluate effects of probiotics on microbiota and surgical recovery. This randomized double-blind trial was performed at four medical centers in South Korea. Of 68 patients expected to undergo anterior sigmoid colon cancer resection, 60 were eligible, of whom 29 and 31 received probiotics and placebo, respectively, for four weeks, starting at one week preoperatively. Third- and/or fourth-week information on anterior resection syndrome (ARS), inflammatory markers, and quality of life was obtained. Stool sample analysis was conducted after randomization and bowel preparation and at three and four postoperative weeks. Bacteria were categorized into Set I (with probiotic effects) and II (colon cancer-associated). The probiotic group's ARS score showed an improving trend (p =0.063), particularly for flatus control (p =0.030). Serum zonulin levels significantly decreased with probiotics. Probiotic ingestion resulted in compositional changes in gut microbiota; greater increases and decreases in Set I and II bacteria, respectively, occurred with probiotics. Compositional increase in Set I bacteria was associated with reduced white blood cells, neutrophils, neutrophil-lymphocyte ratio, and zonulin.Bifidobacteriumcomposition was negatively correlated with zonulin levels in the probiotic group. Probiotics improved postoperative flatus control and modified postoperative changes in microbiota and inflammatory markers.

IFN-gamma, IL-17A, or zonulin rapidly increase the permeability of the blood-brain and small intestinal epithelial barriers: Relevance for neuro-inflammatory diseases

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS

Authors: Rahman, Mohammed T.; Ghosh, Chaitali; Hossain, Mohammed; Linfield, Debra; Rezaee, Fariba; Janigro, Damir; Marchi, Nicola; van Boxel-Dezaire, Anette H. H.

Breakdown of the blood-brain barrier (BBB) precedes lesion formation in the brains of multiple sclerosis (MS) patients. Since recent data implicate disruption of the small intestinal epithelial barrier (IEB) in the pathogenesis of MS, we hypothesized that the increased permeability of the BBB and IEB are mechanistically linked. Zonulin, a protein produced by small intestine epithelium, can rapidly increase small intestinal permeability. Zonulin blood levels are elevated in MS, but it is unknown whether zonulin can also disrupt the BBB. Increased production of IL-17A and IFN-gamma has been implicated in the pathogenesis of MS, epilepsy, and stroke, and these cytokines impact BBB integrity after 24 h. We here report that primary human brain microvascular endothelial cells expressed the EGFR and PAR2 receptors necessary to respond to zonulin, and that zonulin increased BBB permeability to a 40 kDa dextran tracer within 1 h. Moreover, both IL-17A and IFN-gamma also rapidly increased BBB and IEB permeability. By using confocal microscopy, we found that exposure of the IEB to zonulin, IFN-gamma, or IL-17A in vitro rapidly modified the localization of the TJ proteins, ZO-1, claudin-5, and occludin.TJ disassembly was accompanied by marked depolymerization of the penjunctional F-actin cytoskeleton. Our data indicate that IFN-gamma, IL-17A, or zonulin can increase the permeability of the IEB and BBB rapidly in vitro, by modifying TJs and the underlying actin cytoskeleton. These observations may help clarify how the gut-brain axis mediates the pathogenesis of neuro-inflammatory diseases. (C) 2018 Elsevier Inc. All rights reserved.

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