Human ICAM-1/CD54 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 ICAM-1/CD54. 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: ICAM-1/CD54
ICAM-1 (intercellular adhesion molecule-1), also known as CD54, is a transmembrane protein that is upregulated on endothelial and epithelial cells at sites of inflammation. It mediates the vascular adhesion and paracellular migration of leukocytes expressing activated LFA-1 (CD11a/CD18) and Mac-1 (CD11b/CD18). It also binds several non-integrin ligands including CD43/Sialophorin, Fibrinogen, Hyaluronan, rhinoviruses, and Plasmodium falciparum-infected erythrocytes. Soluble ICAM-1 promotes angiogenesis and serves an indicator of vascular endothelial cell activation or damage. Elevated levels of soluble ICAM-1 are associated with cardiovascular disease, type 2 diabetes, organ transplant dysfunction, oxidant stress, increased abdominal fat mass, hypertension, liver disease, certain malignancies, and cerebral malaria.
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 ICAM-1/CD54 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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TNF Induces Laminin-332-Encoding Genes in Endothelial Cells and Laminin-332 Promotes an Atherogenic Endothelial Phenotype
Authors: Hayderi, A;Zegeye, MM;Meydan, S;Sirsjö, A;Kumawat, AK;Ljungberg, LU;
International journal of molecular sciences
Species: Human
Sample Types: Cell Culture Supernates, Cell Lysates
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Are soluble E-selectin, ICAM-1, and VCAM-1 potential predictors for the development of diabetic retinopathy in young adults, 15-34 years of age? A Swedish prospective cohort study
Authors: Ekelund, C;Dereke, J;Nilsson, C;Landin-Olsson, M;
PloS one
Species: Human
Sample Types: Plasma
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Targeting IL-6 trans-signalling by sgp130Fc attenuates severity in SARS-CoV-2 -infected mice and reduces endotheliopathy
Authors: Rodríguez-Hernández, MÁ;Baena-Bustos, M;Carneros, D;Zurita-Palomo, C;Muñoz-Pinillos, P;Millán, J;Padillo, FJ;Smerdou, C;von Kobbe, C;Rose-John, S;Bustos, M;
EBioMedicine
Species: Transgenic Mouse
Sample Types: Serum
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Effects of an Intervention with Selenium and Coenzyme Q10 on Five Selected Age-Related Biomarkers in Elderly Swedes Low in Selenium: Results That Point to an Anti-Ageing Effect-A Sub-Analysis of a Previous Prospective Double-Blind Placebo-Controlled Randomised Clinical Trial
Authors: Alehagen, U;Alexander, J;Aaseth, JO;Larsson, A;Svensson, E;Opstad, TB;
Cells
Species: Human
Sample Types: Plasma
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Impact of Enzymatically Extracted High Molecular Weight Fucoidan on Lipopolysaccharide-Induced Endothelial Activation and Leukocyte Adhesion
Authors: Kirsten, N;Ohmes, J;Mikkelsen, MD;Nguyen, TT;Blümel, M;Wang, F;Tasdemir, D;Seekamp, A;Meyer, AS;Fuchs, S;
Marine drugs
Species: Human
Sample Types: Cell Culture Supernates
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Blood orange juice intake modulates plasma and PBMC microRNA expression in overweight and insulin resistance women: impact on MAPK and NFkappaB signaling pathways
Authors: VC Capetini, BJ Quintanilh, DC de Oliveir, AH Nishioka, LA de Matos, LRP Ferreira, FM Ferreira, GR Sampaio, NMA Hassimotto, FM Lajolo, RA Fock, MM Rogero
The Journal of nutritional biochemistry, 2022-11-26;0(0):109240.
Species: Human
Sample Types: Plasma
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Biomarkers of Exposure and Potential Harm in Exclusive Users of Nicotine Pouches and Current, Former, and Never Smokers: Protocol for a Cross-sectional Clinical Study
Authors: D Azzopardi, LE Haswell, J Frosina, M McEwan, N Gale, J Thissen, F Meichanetz, G Hardie
JMIR research protocols, 2022-10-06;11(10):e39785.
Species: Human
Sample Types: Plasma
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Intestinal Injury Biomarkers Predict Mortality in Pediatric Severe Malaria
Authors: ML Sarangam, R Namazzi, D Datta, C Bond, CPB Vanderpool, RO Opoka, CC John, AL Conroy
MBio, 2022-09-07;0(0):e0132522.
Species: Human
Sample Types: Serum
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Pro-cancerogenic effects of spontaneous and drug-induced senescence of ovarian cancer cells in vitro and in vivo: a comparative analysis
Authors: S Rutecki, P Szulc, M Paku?a, P Uruski, A Radziemski, E Naumowicz, R Moszy?ski, A Tykarski, J Miku?a-Pie, K Ksi??ek
Journal of ovarian research, 2022-07-26;15(1):87.
Species: Human
Sample Types: Cell Culture Supernates
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Depolymerization of fucoidan with endo-fucoidanase changes bioactivity in processes relevant for bone regeneration
Authors: J Ohmes, MD Mikkelsen, TT Nguyen, VHN Tran, S Meier, MS Nielsen, M Ding, A Seekamp, AS Meyer, S Fuchs
Carbohydrate polymers, 2022-02-24;286(0):119286.
Species: Human
Sample Types: Cell Culture Supernates
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Non-HLA Antibodies in Hand Transplant Recipients Are Connected to Multiple Acute Rejection Episodes and Endothelial Activation
Authors: D Sikorska, D Kami?ska, R Catar, M Banasik, H Heidecke, K Schulze-Fo, K Korybalska, R Rutkowski, J ?uczak, J Jab?ecki, A Oko, P Daroszewsk, M Kusztal, W Samborski
Journal of Clinical Medicine, 2022-02-04;11(3):.
Species: Human
Sample Types: Serum
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Clinical and biological markers for predicting ARDS and outcome in septic patients
Authors: J Villar, R Herrán-Mon, E González-H, M Prieto-Gon, A Ambrós, A Rodríguez-, A Muriel-Bom, R Solano, C Cuenca-Rub, A Vidal, C Flores, JM González-M, MI García-Lao, Genetics o
Scientific Reports, 2021-11-22;11(1):22702.
Species: Human
Sample Types: Serum
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Uremic serum damages endothelium by provoking excessive neutrophil extracellular trap formation
Authors: HW Lee, V Nizet, JN An, HS Lee, YR Song, SG Kim, JK Kim
Scientific Reports, 2021-11-02;11(1):21439.
Species: Human
Sample Types: Plasma
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Mimicking of Blood Flow Results in a Distinct Functional Phenotype in Human Non-Adherent Classical Monocytes
Authors: E Wirthgen, M Hornschuh, IM Wrobel, C Manteuffel, J Däbritz
Biology, 2021-08-04;10(8):.
Species: Human
Sample Types: Cell Culture Supernates
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Predictors of Early-Recurrence Atrial Fibrillation after Catheter Ablation in Women and Men with Abnormal Body Weight
Authors: J Budzianows, J Hiczkiewic, K ?ojewska, E Kawka, R Rutkowski, K Korybalska
Journal of Clinical Medicine, 2021-06-18;10(12):.
Species: Human
Sample Types: Serum
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Effects of Bovine Colostrum with or without Egg on In Vitro Bacterial-Induced Intestinal Damage with Relevance for SIBO and Infectious Diarrhea
Authors: RJ Playford, N Choudhry, P Kelly, T Marchbank
Nutrients, 2021-03-22;13(3):.
Species: Human
Sample Types: Cell Lysates
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SIRT7 deficiency suppresses inflammation, induces EndoMT, and increases vascular permeability in primary pulmonary endothelial cells
Authors: AE Wyman, TTT Nguyen, P Karki, ME Tulapurkar, CO Zhang, J Kim, TG Feng, AJ Dabo, NW Todd, IG Luzina, P Geraghty, RF Foronjy, JD Hasday, AA Birukova, SP Atamas, KG Birukov
Sci Rep, 2020-07-27;10(1):12497.
Species: Human
Sample Types: Cell Culture Supernates
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Shiga Toxin Selectively Upregulates Expression of Syndecan-4 and Adhesion Molecule ICAM-1 in Human Glomerular Microvascular Endothelium
Authors: EB Volokhina, WJC Feitz, LM Elders, TJAM van der Ve, NCAJ van de Kar, LPWJ van den He
Toxins (Basel), 2020-07-03;12(7):.
Species: Human
Sample Types: Cell Culture Supernates
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Microinvasion by Streptococcus pneumoniae induces epithelial innate immunity during colonisation at the human mucosal surface
Authors: CM Weight, C Venturini, S Pojar, SP Jochems, J Reiné, E Nikolaou, C Solórzano, M Noursadegh, JS Brown, DM Ferreira, RS Heyderman
Nat Commun, 2019-07-16;10(1):3060.
Species: Human
Sample Types: Cell Culture Supernates
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A Unique Pattern of Mesothelial-Mesenchymal Transition Induced in the Normal Peritoneal Mesothelium by High-Grade Serous Ovarian Cancer
Authors: M Paku?a, P Uruski, A Niklas, A Wo?niak, D Szpurek, A Tykarski, J Miku?a-Pie, K Ksi??ek
Cancers (Basel), 2019-05-13;11(5):.
Species: Human
Sample Types: Cell Culture Supernates
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Systemic inflammation is associated with malaria and preterm birth in women living with HIV on antiretrovirals and co-trimoxazole
Authors: CR McDonald, AM Weckman, AL Conroy, P Olwoch, P Natureeba, MR Kamya, DV Havlir, G Dorsey, KC Kain
Sci Rep, 2019-05-01;9(1):6758.
Species: Human
Sample Types: Plasma
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Blood inflammatory and endothelial markers in women with von Willebrand disease
Authors: I Govorov, K Bremme, A Larsson, M Holmström, E Komlichenk, R Chaireti, M Mints
PLoS ONE, 2019-01-10;14(1):e0210544.
Species: Human
Sample Types: Plasma
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IL-1β-induced ICAM-1 and IL-8 expression/secretion of dental pulp cells is differentially regulated by IRAK and p38
Authors: MC Chang, SI Lin, YH Pan, LD Lin, YL Wang, SY Yeung, HH Chang, JH Jeng
J. Formos. Med. Assoc., 2018-12-14;118(8):1247-1254.
Species: Human
Sample Types: Cell Culture Supernates
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Serum Apolipoprotein E and Other Inflammatory Markers Can Identify Non-Responding Patients to a Dendritic Cell Vaccine
Authors: H Leeman, E Kaminska, D Green, M Bodman-Smi, A Gravett, K Bodman-Smi, J Copier, G Coulton, A Fusi, AG Dalgleish
Transl Oncol, 2018-12-08;12(3):397-403.
Species: Human
Sample Types: Serum
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The immunoproteasome inhibitor ONX-0914 regulates inflammation and expression of contraction associated proteins in myometrium
Authors: S Liong, R Lim, C Nguyen-Ngo, G Barker, HC Parkington, M Lappas
Eur. J. Immunol., 2018-06-28;0(0):.
Species: Human
Sample Types: Tissue Homogenates
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Early-onset preeclampsia predisposes to preclinical diastolic left ventricular dysfunction in the fifth decade of life: An observational study
Authors: A Bokslag, C Franssen, LJ Alma, I Kovacevic, FV Kesteren, PW Teunissen, O Kamp, W Ganzevoort, PL Hordijk, CJM Groot, WJ Paulus
PLoS ONE, 2018-06-12;13(6):e0198908.
Species: Human
Sample Types: Plasma
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Biomarkers of endothelial dysfunction predict sepsis mortality in young infants: a matched�case-control study
Authors: JK Wright, K Hayford, V Tran, GM Al Kibria, A Baqui, A Manajjir, A Mahmud, N Begum, M Siddiquee, KC Kain, A Farzin
BMC Pediatr, 2018-03-23;18(1):118.
Species: Human
Sample Types: Plasma
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Effects of fish and krill oil on gene expression in peripheral blood mononuclear cells and circulating markers of inflammation: a randomised controlled trial
Authors: A Rundblad, KB Holven, I Bruheim, MC Myhrstad, SM Ulven
J Nutr Sci, 2018-03-21;7(0):e10.
Species: Human
Sample Types: Serum
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Serum Soluble Vascular Cell Adhesion Molecule-1 Overexpression Is a Disease Marker in Patients with First-Time Diagnosed Antinuclear Antibodies: A Prospective, Observational Pilot Study
Authors: M Oleszowsky, MF Seidel
Biomed Res Int, 2018-02-01;2018(0):8286067.
Species: Human
Sample Types: Serum
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cfDNA correlates with endothelial damage after cardiac surgery with prolonged cardiopulmonary bypass and amplifies NETosis in an intracellular TLR9-independent manner
Authors: A Paunel-Gör, M Wacker, M El Aita, S Hassan, G Schlachten, A Deppe, YH Choi, E Kuhn, TO Mehler, T Wahlers
Sci Rep, 2017-12-12;7(1):17421.
Species: Human
Sample Types: Plasma
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Pre- and post-surgical evaluation of the inflammatory response in patients with aortic stenosis treated with different types of prosthesis
Authors: ME Soto, JL Salas, J Vargas-Bar, R Marquez, A Rodriguez-, R Bojalil-Pa, I Pérez-Torr, V Guarner-La
BMC Cardiovasc Disord, 2017-04-14;17(1):100.
Species: Human
Sample Types: Serum
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Serum retinol-binding protein-induced endothelial inflammation is mediated through the activation of toll-like receptor 4
Authors: M Du, A Martin, F Hays, J Johnson, RA Farjo, KM Farjo
Mol. Vis., 2017-03-31;23(0):185-197.
Species: Human
Sample Types: Cell Culture Supernates
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Serum Biomarkers of Myocardial Remodeling and Coronary Dysfunction in Early Stages of Hypertrophic Cardiomyopathy in the Young
Authors: E Fernlund, T Gyllenhamm, R Jablonowsk, M Carlsson, A Larsson, J Ärnlöv, P Liuba
Pediatr Cardiol, 2017-03-30;0(0):.
Species: Human
Sample Types: Serum
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Prevention of influenza virus induced bacterial superinfection by standardized Echinacea purpurea, via regulation of surface receptor expression in human bronchial epithelial cells
Authors: V Selvarani, S Roland, S Andy, H James
Virus Res, 2017-03-07;0(0):.
Species: Human
Sample Types: Cell Lysates
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Serum from Varicose Patients Induces Senescence-Related Dysfunction of Vascular Endothelium Generating Local and Systemic Proinflammatory Conditions
Oxid Med Cell Longev, 2016-11-23;2016(0):2069290.
Species: Human
Sample Types: Serum
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ADAM17 substrate release in proximal tubule drives kidney fibrosis
JCI Insight, 2016-08-18;1(13):.
Species: Human
Sample Types: Urine
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Modulation of Endothelial Inflammation by Low and High Magnitude Cyclic Stretch
PLoS ONE, 2016-04-29;11(4):e0153387.
Species: Human
Sample Types: Cell Culture Supernates
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Inflammatory and Angiogenic Factors at Mid-Pregnancy Are Associated with Spontaneous Preterm Birth in a Cohort of Tanzanian Women.
Authors: McDonald C, Darling A, Conroy A, Tran V, Cabrera A, Liles W, Wang M, Aboud S, Urassa W, Fawzi W, Kain K
PLoS ONE, 2015-08-06;10(8):e0134619.
Species: Human
Sample Types: Plasma
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Necroptosis suppresses inflammation via termination of TNF- or LPS-induced cytokine and chemokine production.
Authors: Kearney C, Cullen S, Tynan G, Henry C, Clancy D, Lavelle E, Martin S
Cell Death Differ, 2015-01-23;22(8):1313-27.
Species: Human
Sample Types: Cell Culture Supernates
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Benefits of whole body vibration training in patients hospitalised for COPD exacerbations - a randomized clinical trial.
Authors: Greulich T, Nell C, Koepke J, Fechtel J, Franke M, Schmeck B, Haid D, Apelt S, Filipovic S, Kenn K, Janciauskiene S, Vogelmeier C, Koczulla A
BMC Pulm Med, 2014-04-11;14(0):60.
Species: Human
Sample Types: Serum
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Host biomarkers distinguish dengue from leptospirosis in Colombia: a case-control study.
Authors: Conroy A, Gelvez M, Hawkes M, Rajwans N, Liles W, Villar-Centeno L, Kain K
BMC Infect Dis, 2014-01-20;14(0):35.
Species: Human
Sample Types: Serum
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Induction of pro-inflammatory response of the placental trophoblast by Plasmodium falciparum infected erythrocytes and TNF.
Authors: Vasquez A, Segura C, Blair S
Malar J, 2013-11-15;12(0):421.
Species: Human
Sample Types: Cell Culture Supernates
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Quantification of endothelial microparticles on modified cytometric bead assay and prognosis in chest pain patients.
Authors: Fan, Yuqi, Wang, Lianshen, Li, Yuanmin, Yin, Zhaofang, Xu, Zuojun, Wang, Changqia
Circ J, 2013-11-08;78(1):206-14.
Species: Human
Sample Types: Serum
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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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Endothelium-based biomarkers are associated with cerebral malaria in Malawian children: a retrospective case-control study.
Authors: Conroy AL, Phiri H, Hawkes M, Glover S, Mallewa M, Seydel KB, Taylor TE, Molyneux ME, Kain KC
PLoS ONE, 2010-12-29;5(12):e15291.
Species: Human
Sample Types: Plasma
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Genetic variation of the human urinary tract innate immune response and asymptomatic bacteriuria in women.
Authors: Hawn TR, Scholes D, Wang H, Li SS, Stapleton AE, Janer M, Aderem A, Stamm WE, Zhao LP, Hooton TM
PLoS ONE, 2009-12-15;4(12):e8300.
Species: Human
Sample Types: Urine
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The role of RGD-tagged helical rosette nanotubes in the induction of inflammation and apoptosis in human lung adenocarcinoma cells through the P38 MAPK pathway.
Authors: Suri SS, Rakotondradany F, Myles AJ, Fenniri H, Singh B
Biomaterials, 2009-02-27;30(17):3084-90.
Species: Human
Sample Types: Cell Culture Supernates
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Suspension microarrays for the identification of the response patterns in hyperinflammatory diseases.
Authors: Hsu HY, Wittemann S, Schneider EM, Weiss M, Joos TO
Med Eng Phys, 2008-03-03;30(8):976-83.
Species: Human
Sample Types: Plasma
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Used in Serum and EDTA-Plasma at 1:10 dilution, the kit worked very well, the sample may need further dilution.
We used human serum at a dilution of 1:200. The result was good.