Citrullinated histone h3 clone 11d3 elisa kit

Manufactured by Cayman Chemical

Sourced in United States

The Citrullinated Histone H3 (Clone 11D3) ELISA Kit is a laboratory tool used to detect and quantify citrullinated histone H3, a post-translational modification associated with various biological processes. The kit provides a standardized method for measuring the levels of this target protein in biological samples.

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Lab products found in correlation

Luminex assay, by R&D Systems (1 mentions) Mouse anti human mpo antibody, by Bio-Rad (1 mentions) Human myeloperoxidase quantikine elisa kit, by R&D Systems (1 mentions) Quant it picogreen dsdna assay kit, by Thermo Fisher Scientific (1 mentions) Cytation 5 microplate reader, by Agilent Technologies (1 mentions) Netosis assay kit, by Cayman Chemical (1 mentions) Neutrophil myeloperoxidase activity assay kit, by Cayman Chemical (1 mentions) Multiskan go microplate reader, by Thermo Fisher Scientific (1 mentions) Cell death detection elisaplus kit, by Merck Group (1 mentions)

14 protocols using citrullinated histone h3 clone 11d3 elisa kit

Quantification of NETs-associated MPO-DNA

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NETs-associated MPO-DNA complexes were quantified as previously described.⁷ (link)^,²⁹ Briefly, 5 μg/mL of mouse anti-human MPO antibody (Bio-Rad Antibodies, Hercules, CA) was coated to 96-well microtiter plates. After a 1-hour blocking with 1% bovine serum albumin, serum samples were added together with a peroxidase-labeled anti-DNA monoclonal antibody (component 2 of the Cell Death Detection ELISA PLUS kit, Millipore Sigma/Roche, Oakville, ON). After a 2-hour incubation on the shaker at 320 rpm, the peroxidase substrate was added according to the manufacturer's instructions. The optical absorbance was measured at 405 nm. CRP in serum samples was quantified by nephelometry at the Biochemistry Laboratory at the Montreal Heart Institute. IL-6, MPO, and PTX3 were measured using a Luminex Assay (R&D Systems, Minneapolis, MN). H3Cit was measured using the Citrullinated Histone H3 (Clone 11D3) ELISA Kit (Cayman Chemical, Ann Arbor, MI).

Beaubien-Souligny W., Neagoe P.E., Gagnon D., Denault A.Y, & Sirois M.G. (2019). Increased Circulating Levels of Neutrophil Extracellular Traps During Cardiopulmonary Bypass. CJC Open, 2(2), 39-48.

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Quantifying NETs Biomarkers in Blood

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Serum levels of soluble citrullinated histone H3, a known biomarker of NETosis, were measured in all the studied cases. The citrullinated histone H3 (Clone 11D3) ELISA kit (Cayman Chemical, USA) was used to quantify this biomarker according to the plotted standard curve and results were correlated with d-dimer values (the coagulopathy marker) and levels of surface neutrophilic MPO.

Jamali E., Abbasi M., Tayer A.H., Monfared A.A., Tandel P., Tamaddon G., Kazerooni E.S., Rakhshandehroo S, & Ranjbaran R. (2022). The significance of surface neutrophilic MPO expression level in NETosis and NETosis-associated coagulopathies in covid-19 infected patients. Blood Cells, Molecules & Diseases, 96, 102676.

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Quantification of CitH3 in Hospitalized Patients

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Blood was collected from participants on the second day after hospitalization. 4 ml venous whole blood was anticoagulated with an EDTA tube, gently mixed into a 15 ml centrifuge tube, and centrifugated at 3,000 rpm at 4°C for 20 min. Then the plasma was obtained and stored at −80°C for subsequent use. CitH3 was quantified in plasma using the Citrullinated Histone H3 (clone 11D3) ELISA Kit (Cayman, 501,620) according to the manufacturer’s instructions. All samples were repeated in triplicate. In addition, each patient was inspected for blood routine examination on admission. So we obtain the results directly from the laboratory department.

Li Y., Jiang Q., Zhou X., Wu M., Chen J., Liu H., Dai S., Zheng Z., Zhao X., Zhang C., Shi Z., Zhang H., Gu J., Huang Z., Yin G, & Zhao S. (2022). A prospective marker for the prediction of postoperative deep venous thrombosis: Neutrophil extracellular traps. Frontiers in Cell and Developmental Biology, 10, 1071550.

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Quantification of Plasma Biomarkers

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Plasma levels of cfDNA were measured using a fluorometric assay for double-stranded DNA Quant-iT PicoGreen dsDNA Assay kit (Invitrogen) following the manufacturer’s instructions. Plasma levels of citrullinated histone H3, human myeloperoxidase, neutrophil elastase and VWF were determined by ELISA with Citrullinated Histone H3 (Clone 11D3) ELISA Kit (Cayman Chemicals), Human Myeloperoxidase Quantikine ELISA Kit (R&D Systems), Human PMN Elastase Platinum ELISA (eBioscience, San Diego, CA) or by in-house ELISA using sheep anti-human VWF (Abcam, ab11713) for coating followed by detection with polyclonal rabbit anti-human VWF-HRP (Dako, P0226), respectively.

Perdomo J., Leung H.H., Ahmadi Z., Yan F., Chong J.J., Passam F.H, & Chong B.H. (2019). Neutrophil activation and NETosis are the major drivers of thrombosis in heparin-induced thrombocytopenia. Nature Communications, 10, 1322.

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Citrullinated Histone H3 Quantification

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Citrullinated Histone H3 ELISA was quantified in patient plasma using the Citrullinated Histone H3 (clone 11D3) ELISA kit (Cayman Chemical, 501620) according to manufacturer specifications. Plasma samples (which were previously diluted 1:2 with RPMI) were diluted 1:2 with Assay Buffer. Sample acquisition was performed using the Cytation 5 Microplate Reader (BioTek) at 450nm. The standard curve was fitted with a 4-parameter logistic curve-fitting algorithm using the dr4pl package in R.

LaSalle T.J., Gonye A.L., Freeman S.S., Kaplonek P., Gushterova I., Kays K.R., Manakongtreecheep K., Tantivit J., Rojas-Lopez M., Russo B.C., Sharma N., Thomas M.F., Lavin-Parsons K.M., Lilly B.M., Mckaig B.N., Charland N.C., Khanna H.K., Lodenstein C.L., Margolin J.D., Blaum E.M., Lirofonis P.B., Revach O.Y., Mehta A., Sonny A., Bhattacharyya R.P., Parry B.A., Goldberg M.B., Alter G., Filbin M.R., Villani A.C., Hacohen N, & Sade-Feldman M. (2022). Longitudinal characterization of circulating neutrophils uncovers phenotypes associated with severity in hospitalized COVID-19 patients. Cell Reports Medicine, 3(10), 100779.

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Quantifying H3Cit in NET Formation

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The concentration of H3Cit, which is an integral component of NETs [10 (link)], was assayed using the Citrullinated Histone H3 (Clone 11D3) ELISA Kit (Cayman Chemicals, Ann Arbor, MI, USA). All manufacturer’s instructions were followed.

Lenz M., Maiberger T., Armbrust L., Kiwit A., Von der Wense A., Reinshagen K., Elrod J, & Boettcher M. (2022). cfDNA and DNases: New Biomarkers of Sepsis in Preterm Neonates—A Pilot Study. Cells, 11(2), 192.

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Quantification of Serum Citrullinated Histone H3

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Serum cit-H3 was quantified using the Citrullinated Histone H3 (Clone 11D3) ELISA Kit (Cayman Chemical, Ann Arbor,MI, USA, item no. 501620). This kit contained an anti-histone H3 HRP conjugate, anti-Citrullinated Histone H3 ELISA Strip Plate, Citrullinated Histone ELISA Standard, Immunoassay Buffer B Concentrate (10×), wash buffer concentrate (400×), polysorbate 20, TMB substrate solution, HRP Stop solution and a 96-well cover sheet. A Heidolph shaker (Labotech, Midrand, South Africa) was used. A Nano SPECTROstar plate reader (BMG Labtech, Ortenberg, Germany) interfaced with appropriate software was used to read absorbance and to analyze the data.

Moodley M., Moodley J, & Naicker T. (2024). Placental neutrophil reverse trans-migration and maternal serum neutrophil extracellular trap expression in HIV infection co-morbid pre-eclampsia in women of African ancestry. Histochemistry and cell biology.

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Quantifying Neutrophil Activation and NETosis

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Blood neutrophil (suspended at 10⁶/ml) were stimulated with 5 nM of PMA for 4h. Myeloperoxidase release was assayed using a commercial Neutrophil Myeloperoxidase Activity Assay Kit (Cayman chemical, Cat: 600620). Both soluble Elastase and NET associated elastase were assayed using a NETosis Assay Kit (Cayman chemical, Cat: 601010). Citrullinated H3 (CitH3), which is released from neutrophils during NETosis, was measured by Citrullinated Histone H3 (Clone 11D3) ELISA Kit (Cayman chemical, Cat: 501620).

Schnabel E.L, & Jones A.L. (2001). Isolation and Characterization of Five Erwinia amylovora Bacteriophages and Assessment of Phage Resistance in Strains of Erwinia amylovora. Applied and Environmental Microbiology, 67(1), 59-64.

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Quantifying H3Cit in CSF and Serum

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The measurement of H3Cit in CSF and serum samples was performed with a Citrullinated Histone H3 (Clone 11D3) ELISA Kit (Cayman Chemical; Ann Arbor; Michigan; USA) according to the manufacturers’ instructions in Multiskan Go microplate reader (Thermo Fisher Scientific; Waltham; MA; USA).

Wohlsein J.C., Meurer M., Mörgelin M., Nessler J.N., Flegel T., Schenk H.C., Jurina K., Rentmeister K., Fischer A., Gödde T., Baumgärtner W., von Köckritz-Blickwede M, & Tipold A. (2024). Neutrophil extracellular traps in CSF and serum of dogs with steroid-responsive meningitis-arteritis. PLOS ONE, 19(1), e0295268.

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Evaluation of Inflammatory Markers in Samples

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The following assays were conducted: the General Nicotinamide Adenine Dinucleotide (NAD) ELISA Kit (Ref: MBS2700640-96, MyBioSource, San Diego, California, UnitedStates), to detect the amount of NAD; the Citrullinated Histone H3 (Clone 11D3) ELISA Kit (Ref: 501620, Lot: 0637729, Cayman Chemical Company, Ann Arbor, Michigan, United States), to detect the amount of citrullinated histone H3 (H3Cit); the Cell Death Detection ELISA^PLUS Kit (Ref: 11774425001, Sigma), to detect the amount of nucleosomes; the Porcine IFN gamma (γ) ELISA Kit (Ref: KSC4021, Thermo Fisher Scientific Inc., Waltham, Massachusetts, United States), to detect IFN-γ; and the Porcine IFN alpha (α) ELISA Kit (Ref: ES7RB, Thermo Fisher Scientific Inc.), to detect IFN-α. The assays were performed according to the manufacturer’s recommendations. A sandwich ELISA for the detection of histone–MPO complexes was performed as described before (26 (link)).

Lassnig S., Hennig-Pauka I., Bonilla M.C., Mörgelin M., Imker R., von Köckritz-Blickwede M, & de Buhr N. (2024). Impact of bronchoalveolar lavage from influenza A virus diseased pigs on neutrophil functions and growth of co-infecting pathogenic bacteria. Frontiers in Immunology, 15, 1325269.

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