Human ICAM-1/CD54 Non Allele-specific Quantikine ELISA Kit

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Human ICAM-1/CD54 Non Allele-specific ELISA Standard Curve
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Human ICAM-1/CD54 Non Allele-specific Quantikine ELISA Kit Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
2.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (100 uL), Serum (10 uL), EDTA Plasma (10 uL), Heparin Plasma (10 uL), Urine (50 uL)
Sensitivity
0.053 ng/mL
Assay Range
0.6 - 40 ng/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Urine)
Specificity
Natural and recombinant human ICAM-1
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
Interference
No significant interference observed with available related molecules.

Product Summary

The Quantikine Human ICAM-1/CD54 Non Allele-specific Immunoassay is a 2.5 hour solid-phase ELISA designed to measure human ICAM-1 in cell culture supernates, serum, plasma, and urine. It contains NS0-expressed recombinant human ICAM-1 and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human ICAM-1 showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for naturally occurring human ICAM-1.

Precision

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in twenty separate assays to assess inter-assay precision. Assays were performed by at least three technicians using two lots of components

Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Urine

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean 3.03 9.64 20 3.27 9.81 19.5
Standard Deviation 0.128 0.346 0.973 0.282 0.543 1.22
CV% 4.2 3.6 4.9 8.6 5.5 6.3

Recovery

The recovery of human ICAM-1 spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 103 96-117
Urine (n=4) 94 85-106

Linearity

To assess the linearity of the assay, samples containing and/or spiked with high concentrations of human ICAM-1 were diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human ICAM-1/CD54 Non Allele-specific ELISA Linearity

Data Examples

Human ICAM-1/CD54 Non Allele-specific ELISA Standard Curve

Product Datasheets

Preparation and Storage

Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
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
⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.

Assay Procedure

Refer to the product for complete assay procedure.

Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
  1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
  2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

  3. 100 µL Standard, Control, or Sample
  4.   Add 100 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 1 hour on a horizontal orbital microplate shaker.
  5.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

  6. 200 µL Conjugate
  7.   Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 1 hour on the shaker.
  8.   Aspirate and wash 4 times.

  9. 200 µL Substrate Solution
  10.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.

  11. 50 µL Stop Solution
  12.   Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.

Citation for Human ICAM-1/CD54 Non Allele-specific Quantikine ELISA Kit

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.

1 Citation: Showing 1 - 1

  1. Femoral Artery Atherosclerosis Is Associated With Physical Function Across the Spectrum of the Ankle-Brachial Index: The San Diego Population Study
    Authors: CL Wassel, AM Ellis, NC Suder, E Barinas-Mi, DE Rifkin, NI Forbang, JO Denenberg, AM Marasco, BJ McQuaide, NS Jenny, MA Allison, JH Ix, MH Criqui
    J Am Heart Assoc, 2017;6(7):.
    Species: Human
    Sample Types: Plasma

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