Human chitinase 3 like 1 quantikine elisa kit

Manufactured by R&D Systems

Sourced in United States, United Kingdom

The Human Chitinase 3-like 1 Quantikine ELISA Kit is a quantitative sandwich enzyme-linked immunosorbent assay (ELISA) designed for the measurement of human Chitinase 3-like 1 levels in cell culture supernates, serum, and plasma.

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Spelling variants of this product

Quantikine ELISA kit (1163 citations) Quantikine ELISA (398 citations) ELISAs (144 citations) Quantikine kits (80 citations) Human chitinase-3 quantikine ELISA kit (6 citations) Quantikine ELISA Human Chitinase-3–like 1 (4 citations) Quantikines (4 citations) Quantikine Human (2 citations) Human kits (2 citations)

11 protocols using human chitinase 3 like 1 quantikine elisa kit

CSF Biomarker Measurement Protocol

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CSF samples were collected by LP at the L3/L4 or L4/L5 interspace with a 22gx90mm Sprotte™ needle, by an experienced neurologist. The needle type was chosen to minimize the risk of post-LP headache [5 (link)]. Sampling was performed between 8 and 9 am 3 days apart. A total of 10–12 mL of CSF was collected in polypropylene tubes, centrifuged at 1300g for 10 min, aliquoted and stored in 0.5 mL aliquots at − 80 °C pending analysis within 1 h after sampling.
CSF Aβ38, Aβ40 and Aβ42 concentrations were measured using MSD Abeta Triplex (Meso Scale Discovery, Rockville, Maryland). CSF total tau (T-tau) and phosphorylated tau (P-tau) concentrations were measured using INNOTEST sandwich enzyme-linked immunosorbent assays (ELISAs, Fujirebio, Ghent, Belgium). CSF neurofilament light (NF-L) concentration was measured using the NF-Light ELISA (UmanDiagnostics, Umeå, Sweden). CSF YKL-40 (also called chitinase 3-like 1) concentration was measured using the Human Chitinase 3-like 1 Quantikine ELISA Kit (R&D Systems, Inc. Minneapolis, MN). CSF glial fibrillary acidic protein (GFAP) concentration was measured using an in-house ELISA [6 (link)]. All measurements were performed in one round of experiments with one batch of reagents and baseline and follow-up samples side by side on the assay plates by board-certified laboratory technicians who were blinded to clinical data.

Olsson M., Ärlig J., Hedner J., Blennow K, & Zetterberg H. (2019). Repeated lumbar punctures within 3 days may affect CSF biomarker levels. Fluids and Barriers of the CNS, 16, 37.

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Quantification of Neuroinflammatory Biomarkers

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CSF YKL40 concentrations were measured in duplicate using the MicroVue YKL40 ELISA kit (Quidel Corporation, San Diego, CA) according to the manufacturer's protocol. Brain tissue protein extracts isolated from frontal cortical tissue were measured in duplicate using the Human Chitinase 3-like 1 Quantikine ELISA kit (R&D Systems, Minneapolis, MN) according to manufacture's recommendations. CSF osteopontin concentrations were measured using the human osteopontin ELISA kit (Immuno-Biological Laboratories America, Minneapolis, MN) according to the manufacturer's recommendations. CSF neurofilament light chain concentrations were measured in duplicate using the Human neurofilament, light polypeptide ELISA kit (MyBioSource, Inc., San Diego, CA). Optical density was measured using the ELx800 Absorbance Microplate Reader (BioTek, Winooski, VT).

Bissel S.J., Kofler J., Nyaundi J., Murphey-Corb M., Wisniewski S.R, & Wiley C.A. (2016). Cerebrospinal fluid biomarkers of simian immunodeficiency virus encephalitis. Journal of neuroimmune pharmacology : the official journal of the Society on NeuroImmune Pharmacology, 11(2), 332-347.

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Profiling Soluble Biomarkers in MPS

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Soluble biomarkers in cell culture medium from the MPS were analysed by enzyme-linked immunosorbent assay (ELISA); IL-6 was measured by Human IL-6 DuoSet ELISA (R&D Systems); MCP-1 was measured by Human CCL2/MCP-1 DuoSet ELISA (R&D Systems); pro-collagen 1 was measured by Human Pro-Collagen I α1 DuoSet (R&D Systems); TIMP-1 was measured by Human TIMP-1 Quantikine ELISA Kit (R&D Systems); Fibronectin was measured by Human Fibronectin DuoSet ELISA (R&D Systems); YKL-40 was measured by Human Chitinase 3-like 1 Quantikine ELISA Kit (R&D Systems); Albumin secretion was measured by Human Albumin AssayMax™ ELISA kit (Assay Pro, St. Charles, MO). For all assays, the manufacturer’s protocol was followed.
Cytokine profiles in cell culture medium samples were also analysed by Luminex array for determination of 38 analytes in total, consisting of cytokines, chemokines, and growth factors using the MILLIPLEX MAP Human Cytokine/Chemokine Magnetic Bead Panel - Premixed 38 Plex (Merck Millipore, USA). In total 15 of the analytes were observed to have no detectable expression or had no significant differences between any of the culture conditions so were excluded from the analysis.

Kostrzewski T., Snow S., Battle A.L., Peel S., Ahmad Z., Basak J., Surakala M., Bornot A., Lindgren J., Ryaboshapkina M., Clausen M., Lindén D., Maass C., Young L.M., Corrigan A., Ewart L, & Hughes D. (2021). Modelling human liver fibrosis in the context of non-alcoholic steatohepatitis using a microphysiological system. Communications Biology, 4, 1080.

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Serum Biomarker Measurement Protocol

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Serum periostin concentrations were measured using a proprietary sandwich enzyme-linked immunosorbent assay (Human Periostin/OSF-2 DuoSet ELISA; Shino-test, Kanagawa, Japan), which utilized anti-periostin antibodies (clones SS18A and SS17B)^{41 (link)}. Serum YKL-40 levels were measured in duplicate using a commercial Human Chitinase 3-like 1Quantikine ELISA Kit according to the manufacturer’s instructions (R & D Systems, Inc. Minneapolis, MN, USA).

Lee Y., Lee E., Yon D.K., Jee H.M., Baek H.S., Lee S.W., Cho J.Y, & Han M.Y. (2021). The potential pathways underlying the association of propyl-paraben exposure with aeroallergen sensitization and EASI score using metabolomics analysis. Scientific Reports, 11, 3772.

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Quantifying CSF YKL-40 by ELISA

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CSF YKL-40 concentration was measured using a solid-phase sandwich ELISA (Human Chitinase 3-like 1 Quantikine ELISA kit, R&D Systems, Oxon, UK) according to the manufacturer's protocol: 50 μL of CSF was diluted 2-fold in diluent provided with the kit. Calibrators and controls were prepared according to protocol.
On a capture-antibody precoated 96-well microtiter plate, 50 μL of CSF sample, calibrator, or controls were added followed by addition of 200 μL of conjugate and incubation at room temperature for 2 hours. Finally, each well was washed in the provided washing buffer, followed by addition of 200 μL substrate solution, and subsequently stopped with 50 μL of the provided stop solution. Quantification was carried out at 540/570 nm in a microplate reader.
All biochemical measurements were performed by board-certified laboratory technicians who were blinded to clinical information. The measurements were performed in one round of experiments using one batch of reagents. Intra-assay coefficients of variation were below 10%.

Rostgaard N., Roos P., Portelius E., Blennow K., Zetterberg H., Simonsen A.H, & Nielsen J.E. (2018). CSF neurofilament light concentration is increased in presymptomatic CHMP2B mutation carriers. Neurology, 90(2), e157-e163.

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Cerebrospinal Fluid Biomarker Analysis

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Eight ml of ventricular CSF was collected during the surgical intervention, immediately after shunt insertion and a discard of the first 2 ml. All CSF analyses were performed at the Neurochemistry Laboratory at Sahlgrenska University Hospital, by board-certified laboratory technicians who were blinded to clinical data. Aβ-related biomarkers (Aβ40, Aβ42, sAPPα and sAPPβ) and MCP1 were analyzed by electrochemiluminescence assays (Meso Scale Discovery, Rockville, MD, USA). Validated in-house ELISA methodology was used to analyze NfL [19 (link)), neurogranin [15 (link)), GAP43 [20 (link)) and GFAP [21 (link)), whereas CSF levels of T-tau, and P-tau were measured using commercially available Lumipulse technology (Fujirebio, Ghent, Belgium), as previously described [22 (link)]. YKL-40 was measured using Human Chitinase 3-like 1 Quantikine ELISA Kit (R&D Systems, Minneapolis, MN) [22 (link)]. All concentrations are given in ng/L. All samples were analyzed in one round of experiments using one batch of reagents by board-certified laboratory technicians who were blinded to clinical data. Intra-assay coefficients of variation, monitored using internal quality control samples in the beginning and end of each run, were below 10%.

Grønning R., Jeppsson A., Hellström P., Laurell K., Farahmand D., Zetterberg H., Blennow K., Wikkelsø C, & Tullberg M. (2023). Association between ventricular CSF biomarkers and outcome after shunt surgery in idiopathic normal pressure hydrocephalus. Fluids and Barriers of the CNS, 20, 77.

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CSF Biomarker Analysis Protocol

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CSF samples were obtained by lumbar puncture in the L3/L4 or L4/L5 interspace with patients nonfasting. The samples were collected in polypropylene tubes and gently mixed to avoid gradient effects. All samples were centrifuged within 30 minutes at + 4°C at 2000g for 10 minutes to remove cells and debris and then stored in aliquots at −80°C pending biochemical analysis.
CSF Aβ₄₂, tau, and P‐tau were analyzed using AlzBio3 (Fujirebio, Ghent, Belgium), NFL was analysed using the NF‐light assay (UmanDiagnostics, Umeå, Sweden), YKL‐40 was analyzed using the Human Chitinase 3‐like 1 Quantikine ELISA Kit (R&D, Minneapolis, Minnesota), α‐syn was analyzed using the Covance assay (Covance, Dedham, Massachusetts), and haemoglobin was analyzed with an assay provided by Bethyl Lab, Inc. (Montgomery, Texas). For α‐syn, only samples with haemoglobin <200 ng/ml were used; consequently, 4 samples from PD patients (all PD with long disease duration) and 1 sample from the controls were excluded, all baseline samples.
For P‐tau there were 8 missing in the control group and 16 missing in the PD group. Baseline and follow‐up CSF samples were always analyzed in the same batch. All analyses were performed using 1 batch of reagents by board‐certified laboratory technicians who were blinded to clinical data. Intra‐assay coefficients of variation were below 10%.

Hall S., Surova Y., Öhrfelt A., Blennow K., Zetterberg H, & Hansson O. (2016). Longitudinal Measurements of Cerebrospinal Fluid Biomarkers in Parkinson's Disease. Movement Disorders, 31(6), 898-905.

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Quantifying CSF Biomarkers Using ELISA

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All ELISAs and activity assays were performed in duplicate. Plates were read using a FLUOstar Omega plate reader (BMG LABTECH, UK). Standard curves were fitted with 4‐parameter logistic regression using MARS data analysis software (BMG LABTECH, UK). CSF samples were thawed on ice prior to measurement. Measurements of CSF concentrations and activity were performed using commercially available assays according to the manufacturer’s instructions (CircuLex human Chitotriosidase ELISA, CircuLex human YKL‐39 ELISA, CycLex Chitotriosidase Fluorimetric Assay Kit, MBL, UK; Human Chitinase‐3‐like 1 Quantikine ELISA kit, R&D systems, UK).
Samples were diluted where necessary to achieve a concentration within the linear range of standard curve measurements. Median intraassay and interassay coefficients of variation were below 10% for all assays: CHIT1 intraassay median 6.9% (IQR 2.9%–15.6%), interassay 4.0% (2.4%–5.2%), lower limit of detection (LLOD) 48.3 pg/mL; CHI3L1 5.8% (2.4%–12.09%), 2.6% (1.4%–11.9%), LLOD 3.55 pg/mL; CHI3L2 3.5% (1.2–7.3), 3.0% (0.7%–6.2%), LLOD 35.1 pg/mL; and CHIT1 activity 6.3% (1.7%–13.9%), 1.58% (0.8%–2.5%), LLOD 12.0 nmol/h/mL. Samples for which the analyte value was unmeasurably low were imputed with the LLOD.

Gray E., Thompson A.G., Wuu J., Pelt J., Talbot K., Benatar M, & Turner M.R. (2020). CSF chitinases before and after symptom onset in amyotrophic lateral sclerosis. Annals of Clinical and Translational Neurology, 7(8), 1296-1306.

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Biomarkers of Neurodegenerative Diseases

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Neurofilament light chain (NfL) in the CSF was analysed using the ELISA kit from Uman Diagnostics (UmanDiagnostics AB, Umeå, Sweden) and in the serum with a single-molecule array (Simoa^®, Quanterix Corporation, Lexington, MA, USA). Neurofilament heavy chain (pNfH) was quantified with Phosphorylated Neurofilament H Human ELISA (Biovendor, Heidelberg, Germany). GFAP in the CSF and serum was analysed using Simoa^® (Quanterix Corporation, Lexington, MA, USA). Chitinase-3-like protein 1 (CHI3L1) in the CSF was analysed using the Human Chitinase 3-like 1 Quantikine ELISA kit (R&D Systems, Minneapolis, MN, USA). Aβ1-42 and Tau in the CSF were analysed using Innotest^® and Tau-ELISA, respectively (Fujirebio Germany GmbH, Hanover, Germany). All analyses were performed according to the manufacture’s manual, and samples were run in duplicates. Measurements were interpreted as valid if duplicate measurements showed a coefficient of variation below 15%.

Huss A., Abdelhak A., Mayer B., Tumani H., Müller H.P., Althaus K., Kassubek J., Otto M., Ludolph A.C., Yilmazer-Hanke D, & Neugebauer H. (2022). Association of Serum GFAP with Functional and Neurocognitive Outcome in Sporadic Small Vessel Disease. Biomedicines, 10(8), 1869.

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Quantitative Chitinase 3-like 1 Assay

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CHI3L1 levels were determined using the Human Chitinase 3-like 1 Quantikine ELISA Kit (R&D Systems, Minneapolis, MN, USA). The assay was performed according to the manufacturer's instruction, and CSF was diluted 1:100 and serum 1:50. All samples were measured in duplicates, and intra-assay coefficients of variation (CVs) were <5% and interassay CV was <10%.

Huss A., Otto M., Senel M., Ludolph A.C., Abdelhak A, & Tumani H. (2020). A Score Based on NfL and Glial Markers May Differentiate Between Relapsing–Remitting and Progressive MS Course. Frontiers in Neurology, 11, 608.

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