Human CD163 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 CD163. 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 2 (5 plates): (Catalog # DY008) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.
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 # DY995), or 1% BSA in PBS, pH 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)
Scientific Data
Product Datasheets
Preparation and Storage
Background: CD163
CD163, also known as M130 and p155, is a transmembrane scavenger receptor that is expressed on monocytes and macrophages and is inducible by immunosuppressant glucocorticoids and IL-10. A soluble form is shed from the cell surface by TACE or neutrophil elastase mediated cleavage in response to oxidative stress, Prostaglandin F2a stimulation, or the activation of Fc gamma receptors, TLR1, 2, 5, or 6. CD163 mediates monocyte binding to bacteria, leading to the release of inflammatory cytokines. It is essential for the circulatory clearance of hemoglobin-haptoglobin (Hb-Hp) complexes as well as free hemoglobin. It can also mediate monocyte-erythroblast adhesion and promote erythroblast expansion. CD163 binds and internalizes the cytokine TWEAK, and the ratio of soluble CD163 to TWEAK in the plasma is elevated during atherosclerosis.
Assay Procedure
GENERAL ELISA PROTOCOL
Plate Preparation
- 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.
- 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.
- Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
- 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, 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 CD163 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.
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Citations: Showing 1 - 10
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A Pilot Investigation of the Association Between Vpr Amino Acid Substitutions and Peripheral Immune Marker Levels in People With Human Immunodeficiency Virus: Implications for Neurocognitive Impairment
Authors: Asia, LK;Van Vuren, EJ;Kruger, IM;Williams, ME;
Open forum infectious diseases
Species: Human hepegivirus
Sample Types: Plasma
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The fatal trajectory of pulmonary COVID-19 is driven by lobular ischemia and fibrotic remodelling
Authors: M Ackermann, JC Kamp, C Werlein, CL Walsh, H Stark, V Prade, R Surabattul, WL Wagner, C Disney, AJ Bodey, T Illig, DJ Leeming, MA Karsdal, A Tzankov, P Boor, MP Kühnel, FP Länger, SE Verleden, HM Kvasnicka, HH Kreipe, A Haverich, SM Black, A Walch, P Tafforeau, PD Lee, MM Hoeper, T Welte, B Seeliger, S David, D Schuppan, SJ Mentzer, DD Jonigk
EBioMedicine, 2022-10-04;85(0):104296.
Species: Human
Sample Types: Plasma
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Immune responses in Omicron SARS-CoV-2 breakthrough infection in vaccinated adults
Authors: H Kared, AS Wolf, A Alirezayla, A Ravussin, G Solum, TT Tran, F Lund-Johan, JT Vaage, LS Nissen-Mey, UC Nygaard, O Hungnes, AH Robertson, LM Næss, L Trogstad, P Magnus, LA Munthe, S Mjaaland
Nature Communications, 2022-07-18;13(1):4165.
Species: Human
Sample Types: Plasma
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The performance of soluble CD163 as a non-invasive biomarker of liver damage in chronically HCV and HCV/HIV infected subjects
Authors: V Cairoli, E De Matteo, P Casciato, B Ameigeiras, MV Preciado, P Valva
PLoS ONE, 2022-07-07;17(7):e0270911.
Species: Human
Sample Types: Plasma
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Arginase 1 is a marker of protection against illness in contacts of leprosy patients
Authors: RB da Silva P, MA Mendes, VC Soares, J França-Cos, AM Sales, NC Duppré, V de Matos B, TP da Silva, PT Bozza, MT Bozza, EN Sarno, MO Moraes, GM Sperandio, RO Pinheiro
Scientific Reports, 2022-05-12;12(1):7850.
Species: Human
Sample Types: Serum
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A 3D Microfluidic ELISA for the Detection of Severe Dengue: Sensitivity Improvement and Vroman Effect Amelioration by EDC-NHS Surface Modification
Authors: H Maeno, PF Wong, S AbuBakar, M Yang, SS Sam, J Jamil-Abd, A Shunmugara, M Mustafa, RM Said, E Mageswaren, A Azmel, A Mat Jelani
Micromachines, 2021-11-30;12(12):.
Species: Human
Sample Types: Serum
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Inflammatory pathways amongst people living with HIV in Malawi differ according to socioeconomic status
Authors: C Kelly, W Tinago, D Alber, P Hunter, N Luckhurst, J Connolly, F Arrigoni, A Garcia Abn, R Kamn'gona, I Sheha, M Chammudzi, K Jambo, J Mallewa, A Rapala, PWG Mallon, H Mwandumba, N Klein, S Khoo
PLoS ONE, 2021-08-25;16(8):e0256576.
Species: Human
Sample Types: Plasma
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Exploring the potential effect of paricalcitol on markers of inflammation in de novo renal transplant recipients
Authors: HK Pihlstrøm, T Ueland, AE Michelsen, P Aukrust, F Gatti, C Hammarströ, M Kasprzycka, J Wang, G Haraldsen, G Mjøen, DO Dahle, K Midtvedt, IA Eide, A Hartmann, H Holdaas
PLoS ONE, 2020-12-16;15(12):e0243759.
Species: Human
Sample Types: Plasma
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Persistence of monocyte activation under treatment in people followed since acute HIV-1 infection relative to participants at high or low risk of HIV infection
Authors: S Novelli, C Lécuroux, C Goujard, J Reynes, A Villemant, L Blum, A Essat, V Avettand-F, O Launay, JM Molina, C Bourgeois, L Meyer
EBioMedicine, 2020-11-26;62(0):103129.
Species: Human
Sample Types: Plasma
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HIV-associated gut dysbiosis is independent of sexual practice and correlates with noncommunicable diseases
Authors: I Vujkovic-C, O Sortino, E Verheij, J Sklar, FW Wit, NA Kootstra, B Sellers, JM Brenchley, J Ananworani, MSV Loeff, Y Belkaid, P Reiss, I Sereti
Nat Commun, 2020-05-15;11(1):2448.
Species: Human
Sample Types: Plasma
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Biofluid quantification of TWEAK/Fn14 axis in combination with a selected biomarker panel improves assessment of prostate cancer aggressiveness
Authors: X Ruiz-Plaza, E Rodríguez-, M Alves, A Altuna-Coy, J Lozano-Bar, M Portero-Ot, JF García-Fon, S Martínez-G, J Segarra, MR Chacón
J Transl Med, 2019-09-09;17(1):307.
Species: Human
Sample Types: Serum
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Soluble CD163 and TWEAK in early pregnancy gestational diabetes and later glucose intolerance
Authors: J Dereke, J Nilsson, C Nilsson, H Strevens, M Landin-Ols, M Hillman
PLoS ONE, 2019-05-09;14(5):e0216728.
Species: Human
Sample Types: Plasma
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Prevalence and Outcome of Secondary Hemophagocytic Lymphohistiocytosis Among SIRS Patients: Results from a Prospective Cohort Study
Authors: GA Gualdoni, GA Hofmann, P Wohlfarth, HM Winkler, S Winkler, H Haslacher, R Thalhammer, A Makristath, F Ratzinger, H Burgmann
J Clin Med, 2019-04-19;8(4):.
Species: Human
Sample Types: Plasma
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Urinary and serum soluble CD25 complements urinary soluble CD163 to detect active renal anti-neutrophil cytoplasmic autoantibody-associated vasculitis: a cohort study
Authors: GJ Dekkema, WH Abdulahad, T Bijma, SM Moran, L Ryan, MA Little, CA Stegeman, P Heeringa, JF Sanders
Nephrol. Dial. Transplant., 2019-02-01;0(0):.
Species: Human
Sample Types: Serum
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Biomarkers and immunoprofiles associated with fetal abnormalities of ZIKV-positive pregnancies
Authors: SS Foo, W Chen, Y Chan, WS Lee, SA Lee, G Cheng, K Nielsen-Sa, P Brasil, JU Jung
JCI Insight, 2018-11-02;3(21):.
Species: Human
Sample Types: Serum
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Epigenetic alterations are associated with monocyte immune dysfunctions in HIV-1 infection
Authors: MS Espíndola, LS Soares, LJ Galvão-Lim, FA Zambuzi, MC Cacemiro, VS Brauer, CM Marzocchi-, M de Souza G, LR Amaral, OA Martins-Fi, VR Bollela, FG Frantz
Sci Rep, 2018-04-03;8(1):5505.
Species: Human
Sample Types: Plasma
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Impact of early cART on HIV blood and semen compartments at the time of primary infection
Authors: A Chéret, C Durier, A Mélard, M Ploquin, J Heitzmann, C Lécuroux, V Avettand-F, L David, G Pialoux, JM Chennebaul, M Müller-Tru, C Goujard, C Rouzioux, L Meyer
PLoS ONE, 2017-07-14;12(7):e0180191.
Species: Human
Sample Types: Plasma
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Monocyte/macrophage and T cell activation markers are not independently associated with MI risk in healthy individuals - results from the HUNT Study
Authors: T Ueland, LE Laugsand, LJ Vatten, I Janszky, C Platou, AE Michelsen, JK Damås, P Aukrust, BO Åsvold
Int. J. Cardiol., 2017-06-12;0(0):.
Species: Human
Sample Types: Serum
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High Cellular Monocyte Activation in People Living With Human Immunodeficiency Virus on Combination Antiretroviral Therapy and Lifestyle-Matched Controls Is Associated With Greater Inflammation in Cerebrospinal Fluid
Authors: T Booiman, FW Wit, I Maurer, D De Frances, CA Sabin, AM Harskamp, M Prins, P Garagnani, C Pirazzini, C Franceschi, D Fuchs, M Gisslén, A Winston, P Reiss, NA Kootstra
Open Forum Infect Dis, 2017-05-25;4(3):ofx108.
Species: Human
Sample Types: Plasma
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Soluble plasma proteins ST2 and CD163 as early biomarkers of nephropathy in Swedish patients with diabetes, 15-34�years of age: a prospective cohort study
Authors: M Samuelsson, J Dereke, MK Svensson, M Landin-Ols, M Hillman, on the beh
Diabetol Metab Syndr, 2017-05-25;9(0):41.
Species: Human
Sample Types: Plasma
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Identification of promising plasma immune biomarkers to differentiate active pulmonary tuberculosis
Cytokine, 2016-08-31;88(0):99-107.
Species: Human
Sample Types: Plasma
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Elevated Basal Pre-infection CXCL10 in Plasma and in the Small Intestine after Infection Are Associated with More Rapid HIV/SIV Disease Onset
PLoS Pathog, 2016-08-10;12(8):e1005774.
Species: Human
Sample Types: Plasma
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The Human Endogenous Protection System against Cell-Free Hemoglobin and Heme Is Overwhelmed in Preeclampsia and Provides Potential Biomarkers and Clinical Indicators.
Authors: Gram M, Anderson U, Johansson M, Edstrom-Hagerwall A, Larsson I, Jalmby M, Hansson S, Akerstrom B
PLoS ONE, 2015-09-14;10(9):e0138111.
Species: Human
Sample Types: Plasma
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Serum sCD163 levels are associated with type 2 diabetes mellitus and are influenced by coffee and wine consumption: results of the Di@bet.es study.
Authors: Rojo-Martinez G, Maymo-Masip E, Rodriguez M, Solano E, Goday A, Soriguer F, Valdes S, Chaves F, Delgado E, Colomo N, Hernandez P, Vendrell J, Chacon M
PLoS ONE, 2014-06-30;9(6):e101250.
Species: Human
Sample Types: Serum
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We have been using this kit for almost 2 years. I would say this an excellent kit for serum samples. It has a user-friendly protocol which allows researchers to perform assay without any interruptions.