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

  
  • 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.
Control Available
QC236, Quantikine Immunoassay Control Group 236 - Please Inquire
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
Sample123123
n202020202020
Mean3.039.64203.279.8119.5
Standard Deviation0.1280.3460.9730.2820.5431.22
CV%4.23.64.98.65.56.3

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 Quantikine ELISA Kit
Preparation and Storage
  • 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;
    Related Research Areas
    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.
    ELISA Controls
    Description Application Cat# Citations Images  

    Quantikine Immunoassay Control Group 236

    QC236
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    Supplemental ELISA Products
    Description Application Cat# Citations Images  

    Quantikine ELISA Wash Buffer 1

    ELISA WA126 6
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