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Human ICAM1 ELISA Kit, Fluorescent (ab229383)

Key features and details

  • One-wash 90 minute protocol
  • Sensitivity: 2.4 pg/ml
  • Range: 4.9 pg/ml - 20000 pg/ml
  • Sample type: Adherent cells, Cell culture extracts, Cell culture supernatant, Cit plasma, EDTA Plasma, Hep Plasma, Serum, Tissue Extracts
  • Detection method: Fluorescent
  • Assay type: Sandwich (quantitative)
  • Reacts with: Human

Overview

  • Product name

    Human ICAM1 ELISA Kit, Fluorescent
    See all ICAM1 kits

  • Detection method

    Fluorescent

  • Precision

    Intra-assay
    Sample n Mean SD CV%
    General 6 6%
    Inter-assay
    Sample n Mean SD CV%
    General 24 7%

  • Sample type

    Cell culture supernatant, Serum, Cell culture extracts, Adherent cells, Tissue Extracts, Hep Plasma, EDTA Plasma, Cit plasma

  • Assay type

    Sandwich (quantitative)

  • Sensitivity

    2.4 pg/ml

  • Range

    4.9 pg/ml - 20000 pg/ml

  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Serum 103 99% - 110%
    Plasma 113 96% - 126%
    Cell culture media 98 91% - 102%
    Extraction Buffer 87 82% - 97%

  • Assay time

    1h 30m

  • Assay duration

    One step assay

  • Species reactivity

    Reacts with: Human
    Does not react with: Goat, Cow, Pig

  • Product overview

    ICAM1 (CD54) in vitro CatchPoint SimpleStep ELISA (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of ICAM1 (CD54) protein in human cell culture supernatant, plasma, serum, and cell extracts.


    This CatchPoint SimpleStep ELISA kit has been optimized for Molecular Devices Microplate Readers. Click here for a list of recommended Microplate Readers.
    If using a Molecular Devices’ plate reader supported by SoftMax® Pro software, a preconfigured protocol for these CatchPoint SimpleStep ELISA Kits is available with all the protocol and analysis settings at www.softmaxpro.org


    The CatchPoint SimpleStep ELISA employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material.  CatchPoint HRP Development Solution containing the Stoplight Red Substrate is added. During incubation, the substrate is catalyzed by HRP generating a fluorescent product. Signal is generated proportionally to the amount of bound analyte and the intensity is measured in a fluorescence plater reader at 530/570/590 nm Excitation/Cutoff/Emission.

  • Notes

    ICAM1, is a cell surface glycoprotein typically expressed in endothelial cells and cells of the immune system.  The extracellular portion of ICAM-1 forms five immunoglobulin like domains attached to a single hydrophobic transmembrane region and a short cytoplasmic tail.  ICAM-1, binds to the Leukocyte Integrins LFA-1 (CD11a/CD18) and Mac-1 (CD11b/CD18) as well as to non-integrin ligands such as CD43, fibrinogen, hyaluronan, Rhinoviruses and Plasmodium falciparum-infected erythrocytes.  Binding to LFA-1 facilitates trans-endothelial leukocyte migration to areas of inflammation via promotion of endothelial apical cups assembly.

    Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
    It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

  • Platform

    Pre-coated microplate (12 x 8 well strips)

Properties

Images

  • Other - Human ICAM1 ELISA Kit, Fluorescent (ab229383)
    Other - Human ICAM1 ELISA Kit, Fluorescent (ab229383)

    SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.

     

  • Example of human ICAM1 (CD54) standard curve in Sample Diluent NS.
    Example of human ICAM1 (CD54) standard curve in Sample Diluent NS.

    Background-subtracted data values (mean +/- SD) are graphed.

  • Specificity of ICAM-1 signal on stimulated and unstimulated media supernatants.
    Specificity of ICAM-1 signal on stimulated and unstimulated media supernatants.

    Human PBMCs were cultured in RPMI supplemented with 10% fetal calf serum, 2 mM L-glutamine, 100 U/mL penicillin, and 100 µg/mL streptomycin. Cells were cultured for 2 days at 37⁰C in the presence or absence of PHA. The concentrations of ICAM-1 were interpolated from the calibration curve and corrected for sample dilution. The mean ICAM-1 concentration was determined to be 320 pg/mL in unstimulated PBMC supernatants and 2,654 pg/mL in stimulated PBMC supernatants.

  • Frequency histogram of ICAM-1 levels in serum of individual normal healthy donors.
    Frequency histogram of ICAM-1 levels in serum of individual normal healthy donors.

    The levels of ICAM-1 in serum samples were tested from ten individual healthy donors. Levels were interpolated from the standard curve and corrected for sample dilution. The levels of ICAM-1 are shown for the percentage of individuals within each 100 ng/mL bin center of the distribution. The mean level of ICAM-1 was 469 ng/mL with a range of 347 to 629 ng/mL and a standard deviation of 77 ng/mL.

  • Example of ICAM1 dynamic range in Raji cell extracts.
    Example of ICAM1 dynamic range in Raji cell extracts.

    The curve was prepared by loading 5 µg/mL of Raji cell extracts, followed by a 1:2 titration series. Background-subtracted data values (mean +/- SD) are graphed.

  • Comparison of ICAM-1 levels in three human cell culture lysates.
    Comparison of ICAM-1 levels in three human cell culture lysates.

    The levels of ICAM-1 protein were assessed in three human cell line lysates loaded at 2.5 µg/mL of protein. The raw OD 450nm signal for each sample was interpolated from an ICAM1 standard curve.

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