Human TREM-1 DuoSet ELISA Summary
* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.
This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human TREM-1. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.
Product Features
- Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
- Development protocols are provided to guide further assay optimization
- Assay can be customized to your specific needs
- Economical alternative to complete kits
Kit Content
- Capture Antibody
- Detection Antibody
- Recombinant Standard
- Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)
Other Reagents Required
DuoSet Ancillary Reagent Kit 3 (5 plates): (Catalog # DY009) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 3.
Normal Goat Serum: (Catalog # DY005)
The components listed above may be purchased separately:
PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered
Wash Buffer: (Catalog # WA126), or 0.05% Tween® 20 in PBS, pH 7.2-7.4
Reagent Diluent: (Catalog # DY004), or 5% Tween 20 in PBS, 7.2-7.4, 0.2 μm filtered
Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)
Stop Solution: 2 N H2SO4 (Catalog # DY994)
Microplates: R&D Systems (Catalog # DY990)
Plate Sealers: ELISA Plate Sealers (Catalog # DY992)
Normal Goat Serum: (Catalog # DY005)
Scientific Data
Product Datasheets
Preparation and Storage
Background: TREM-1
The TREM family of receptors regulates the activity of various cell types of the immune system including neutrophils, monocyte/macrophages, microglia, and dendritic cells. The receptors are members of the immunoglobulin superfamily, characterized structurally by a single V-set immunoglobulin (Ig) domain and a short cytoplasmic tail. TREMs associate with the DAP12 signal adapter, an immunoreceptor tyrosine-based activation motif- (ITAM)-containing protein that is phosphorylated and transduces signals when activated.
Assay Procedure
GENERAL ELISA PROTOCOL
Plate Preparation
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Dilute the Capture Antibody to the working concentration in PBS without carrier protein. Immediately coat a 96-well microplate with 100 μL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
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Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 μL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
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Block plates by adding 300 μL of Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
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Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.
Assay Procedure
- Add 100 μL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
- Repeat the aspiration/wash as in step 2 of Plate Preparation.
- Add 100 μL of the Detection Antibody, diluted in Reagent Diluent with NGS, to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
- Repeat the aspiration/wash as in step 2 of Plate Preparation.
- Add 100 μL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
- Repeat the aspiration/wash as in step 2.
- Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
- Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
- Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.
Citations for Human TREM-1 DuoSet ELISA
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
17
Citations: Showing 1 - 10
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Influence of trem-1 gene polymorphisms on cytokine levels during malaria by Plasmodium vivax in a frontier area of the Brazilian Amazon
Authors: de Jesus, MCS;Cerilo-Filho, M;Ramirez, ADR;Menezes, RAO;Gomes, MSM;Cassiano, GC;Gurgel, RQ;Silva, JRS;Moura, TR;Pratt-Riccio, LR;Baptista, ARS;Storti-Melo, LM;Machado, RLD;
Cytokine
Species: Human
Sample Types: Serum
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The Interplay among Glucocorticoid Therapy, Platelet-Activating Factor and Endocannabinoid Release Influences the Inflammatory Response to COVID-19
Authors: JCS de Carvalh, PV da Silva-N, DM Toro, CA Fuzo, V Nardini, VE Pimentel, MM Pérez, TFC Fraga-Silv, CNS Oliveira, AM Degiovani, FM Ostini, MR Feitosa, RS Parra, JJR da Rocha, O Feres, FC Vilar, GG Gaspar, IKFM Santos, APM Fernandes, SR Maruyama, EMS Russo, VLD Bonato, CRB Cardoso, M Dias-Baruf, LH Faccioli, CA Sorgi, On Behalf
Viruses, 2023-02-19;15(2):.
Species: Human
Sample Types: Plasma
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Circulating sTREM-1 as a predictive biomarker of pediatric multisystemic inflammatory syndrome (MIS-C)
Authors: GS Gonçalves, S Correa-Sil, Y Zheng, I Avelar, MM Montenegro, AEF Ferreira, V Bain, TT Fink, P Suguita, C Astley, L Lindoso, F Martins, OM Matsuo, JCOA Ferreira, I Firigato, F de Toledo, M Fernanda B, C Artur A da, M Carneiro-S, HHS Marques, P Palmeira
Cytokine, 2022-11-18;161(0):156084.
Species: Human
Sample Types: Serum
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Soluble triggering receptor expressed on myeloid cells-1 (sTREM-1) and other inflammatory mediators in malaria by Plasmodium vivax during enteroparasites coinfection
Authors: MCS de Jesus, JHR Barbosa, RAO Menezes, MDSM Gomes, LGS Bomfim, TS Pimenta, ARS Baptista, RLD Machado, TR de Moura, LM Storti-Mel
PLoS ONE, 2022-06-24;17(6):e0270007.
Species: Human
Sample Types: Serum
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sTREM-1 Predicts Disease Severity and Mortality in COVID-19 Patients: Involvement of Peripheral Blood Leukocytes and MMP-8 Activity
Authors: PV da Silva-N, JCS de Carvalh, VE Pimentel, MM Pérez, DM Toro, TFC Fraga-Silv, CA Fuzo, CNS Oliveira, LC Rodrigues, JGM Argolo, I Carmona-Ga, NT Neto, COS Souza, TM Fernandes, VAF Bastos, AM Degiovani, LF Constant, FM Ostini, MR Feitosa, RS Parra, FC Vilar, GG Gaspar, JJR da Rocha, O Feres, FG Frantz, RF Gerlach, SR Maruyama, EMS Russo, AL Viana, APM Fernandes, IKFM Santos, VLD Bonato, AL Boechat, A Malheiro, RT Sadikot, M Dias-Baruf, CRB Cardoso, LH Faccioli, CA Sorgi, On Behalf
Viruses, 2021-12-15;13(12):.
Species: Human
Sample Types: Plasma
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sTREM-1 assessment as a diagnostic biomarker for suspected acute appendicitis in children
Authors: TT Klein, E Kohn, B Klin, T Ziv-Baran, E Kozer, M Berkovitch, I Abu Kishk
Asian journal of surgery, 2021-03-23;0(0):.
Species: Human
Sample Types: Urine
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Elevated Plasma Soluble Triggering Receptor Expressed on Myeloid Cells-1 Level in Patients with Acute Coronary Syndrome (ACS): A Biomarker of Disease Severity and Outcome
Authors: S Shiber, V Kliminski, K Orvin, I Sagy, M Vaturi, R Kornowski, M Drescher, Y Molad
Mediators of Inflammation, 2021-03-15;2021(0):8872686.
Species: Human
Sample Types: Plasma
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Elevated serum TREM-1 is associated with periodontitis and disease activity in rheumatoid arthritis
Authors: N Inanc, G Mumcu, M Can, M Yay, A Silbereise, D Manoil, H Direskenel, N Bostanci
Scientific Reports, 2021-02-03;11(1):2888.
Species: Human
Sample Types: Serum
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Salivary Biomarkers of Oral Inflammation Are Associated With Cardiovascular Events and Death Among Kidney Transplant Patients
Authors: F Ortiz, KM Nylund, H Ruokonen, JH Meurman, J Furuholm, N Bostanci, T Sorsa
Transplant. Proc., 2020-08-04;0(0):.
Species: Human
Sample Types: Saliva
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Diagnostic value of serum soluble triggering expressed receptor on myeloid cells 1 (sTREM-1) in suspected sepsis: a meta-analysis
Authors: W Chang, F Peng, SS Meng, JY Xu, Y Yang
BMC Immunol., 2020-01-13;21(1):2.
Species: Human
Sample Types: Serum
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Oxidized Low-Density Lipoprotein (Ox-LDL) and Triggering Receptor-Expressed Myeloid Cell (TREM-1) Levels Are Associated with Cardiometabolic Risk in Nonobese, Clinically Healthy, and Young Adults
Authors: CMP Vázquez, JO Costa, LGS Bomfim, LV Pires, DG da Silva, KF Fukutani, AR de Jesus, N de Jesus S, G de Jesus S, TR de Moura, K Barbosa
Oxid Med Cell Longev, 2019-02-28;2019(0):7306867.
Species: Human
Sample Types: Serum
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Elevated plasma level of soluble triggering receptor expressed on myeloid cells-1 is associated with inflammation activity and is a potential biomarker of thrombosis in primary antiphospholipid syndrome
Authors: Y Edel, V Kliminski, E Pokroy-Sha, S Oren, A Dortort La, Y Pri-Paz Ba, M Egbaria, Y Molad
Arthritis Res. Ther., 2019-01-07;21(1):10.
Species: Human
Sample Types: Plasma
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Pleiotropy of ABO gene: correlation of rs644234 with E-selectin and lipid levels
Authors: AA Arguinano, NC Ndiaye, C Masson, S Visvikis-S
Clin. Chem. Lab. Med., 2018-04-25;0(0):.
Species: Human
Sample Types:
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In vivo attenuation and genetic evolution of a ST247-SCCmecI MRSA clone after 13 years of pathogenic bronchopulmonary colonization in a patient with cystic fibrosis: implications of the innate immune response.
Authors: Lopez-Collazo E, Jurado T, de Dios Caballero J, Perez-Vazquez M, Vindel A, Hernandez-Jimenez E, Tamames J, Cubillos-Zapata C, Manrique M, Tobes R, Maiz L, Canton R, Baquero F, Del Campo R
Mucosal Immunol, 2014-08-13;8(2):362-71.
Species: Human
Sample Types: Cell Culture Supernates
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Role of MMPs in orchestrating inflammatory response in human monocytes via a TREM-1-PI3K-NF-kappaB pathway.
J. Cell. Sci., 2012-03-29;91(6):933-45.
Species: Human
Sample Types: Cell Culture Supernates
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Combinations of Host Biomarkers Predict Mortality among Ugandan Children with Severe Malaria: A Retrospective Case-Control Study.
Authors: Erdman LK, Dhabangi A, Musoke C, Conroy AL, Hawkes M, Higgins S, Rajwans N, Wolofsky KT, Streiner DL, Liles WC, Cserti-Gazdewich CM, Kain KC
PLoS ONE, 2011-02-25;6(2):e17440.
Species: Human
Sample Types: Plasma
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Metalloproteinases shed TREM-1 ectodomain from lipopolysaccharide-stimulated human monocytes.
Authors: Gomez-Pina V, Soares-Schanoski A, Rodriguez-Rojas A, Del Fresno C, Garcia F, Vallejo-Cremades MT, Fernandez-Ruiz I, Arnalich F, Fuentes-Prior P, Lopez-Collazo E
J. Immunol., 2007-09-15;179(6):4065-73.
Species: Human
Sample Types: Cell Culture Supernates
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