Human ICAM-1/CD54 DuoSet ELISA

Catalog # Availability Size / Price Qty
DY720
DY720-05
Ancillary Products Available
Human ICAM-1 / CD54 ELISA Standard Curve
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Product Details
Procedure
Citations (22)
FAQs
Supplemental Products
Reviews (3)

Human ICAM-1/CD54 DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
Sufficient Materials
For five or fifteen 96-well plates*
Specificity
Please see the product datasheet

* 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)

Data Example

Human ICAM-1 / CD54 ELISA Standard Curve

Product Datasheets

Preparation and Storage

Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

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.

Long Name:
Intercellular Adhesion Molecule 1
Entrez Gene IDs:
3383 (Human); 15894 (Mouse); 25464 (Rat)
Alternate Names:
BB2; CD54 antigen; CD54; cell surface glycoprotein P3.58; human rhinovirus receptor; ICAM1; ICAM-1; intercellular adhesion molecule 1 (CD54), human rhinovirus receptor; intercellular adhesion molecule 1; Major group rhinovirus receptor; P3.58

Assay Procedure

GENERAL ELISA PROTOCOL

Plate Preparation

  1. 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.
  2. 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.
  3. Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
  4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

Assay Procedure

  1. 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.
  2. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  3. 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.
  4. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  5. 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.
  6. Repeat the aspiration/wash as in step 2.
  7. Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  8. Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  9. 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.

22 Citations: Showing 1 - 10
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  1. 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;0(0):.
    Species: Human
    Sample Types: Tissue Supernates
  2. 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;13(6):e0198908.
    Species: Human
    Sample Types: Plasma
  3. 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;2018(0):8286067.
    Species: Human
    Sample Types: Serum
  4. 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;18(1):118.
    Species: Human
    Sample Types: Plasma
  5. 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;7(1):17421.
    Species: Human
    Sample Types: Plasma
  6. 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;0(0):.
    Species: Human
    Sample Types: Serum
  7. 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;17(1):100.
    Species: Human
    Sample Types: Serum
  8. 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;23(0):185-197.
    Species: Human
    Sample Types: Cell Culture Supernates
  9. Modulation of Endothelial Inflammation by Low and High Magnitude Cyclic Stretch
    PLoS ONE, 2016;11(4):e0153387.
    Species: Human
    Sample Types: Cell Culture Supernates
  10. ADAM17 substrate release in proximal tubule drives kidney fibrosis
    JCI Insight, 2016;1(13):.
    Species: Human
    Sample Types: Urine
  11. Serum from Varicose Patients Induces Senescence-Related Dysfunction of Vascular Endothelium Generating Local and Systemic Proinflammatory Conditions
    Oxid Med Cell Longev, 2016;2016(0):2069290.
    Species: Human
    Sample Types: Serum
  12. 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;22(8):1313-27.
    Species: Human
    Sample Types: Cell Culture Supernates
  13. 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;10(8):e0134619.
    Species: Human
    Sample Types: Plasma
  14. 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;14(0):35.
    Species: Human
    Sample Types: Serum
  15. 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, 2014;78(1):206-14.
    Species: Human
    Sample Types: Serum
  16. 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;14(0):60.
    Species: Human
    Sample Types: Serum
  17. 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;12(0):421.
    Species: Human
    Sample Types: Cell Culture Supernates
  18. 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;6(2):e17440.
    Species: Human
    Sample Types: Plasma
  19. 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;5(12):e15291.
    Species: Human
    Sample Types: Plasma
  20. 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;30(17):3084-90.
    Species: Human
    Sample Types: Cell Culture Supernates
  21. 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;4(12):e8300.
    Species: Human
    Sample Types: Urine
  22. 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;30(8):976-83.
    Species: Human
    Sample Types: Plasma

FAQs

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Reviews for Human ICAM-1/CD54 DuoSet ELISA

Average Rating: 3.7 (Based on 3 Reviews)

5 Star
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Human ICAM-1/CD54 DuoSet ELISA
By Anonymous on 10/09/2018
Sample Tested: EDTA Plasma

Human ICAM-1/CD54 DuoSet ELISA
By Anonymous on 08/28/2017
Sample Tested: Serum

We used human serum at a dilution of 1:200. The result was good.


Human ICAM-1/CD54 DuoSet ELISA
By Anonymous on 12/07/2016
Sample Tested: EDTA Plasma

We run human serum and plasma samples at a 1:200 dilution in this assay. The assay had a poor precision with a high inter- and intra-coefficient of variation.