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, Heparin Plasma, EDTA Plasma, Citrate 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: Mouse, Rat, Rabbit, Goat, Guinea pig, Hamster, Cow, Dog, 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.

  • Tested applications
    Suitable for: Sandwich ELISAmore details
  • Platform
    Pre-coated microplate (12 x 8 well strips)

Properties

Applications

Our Abpromise guarantee covers the use of ab229383 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Sandwich ELISA Use at an assay dependent concentration.

Images

  • ELISA Protocol Summary
  • Background-subtracted data values (mean +/- SD) are graphed.

  • 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.

  • 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.

  • 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.

  • 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.

Protocols

References

ab229383 has not yet been referenced specifically in any publications.

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