Overview

  • Product name

    Human CD14 ELISA Kit
    See all CD14 kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Human serum 8 2.8%
    Inter-assay
    Sample n Mean SD CV%
    Human serum 3 2.1%
  • Sample type

    Cell culture supernatant, Serum, Heparin Plasma, EDTA Plasma, Citrate Plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    4 pg/ml
  • Range

    31.3 pg/ml - 2000 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Cell culture supernatant 102 95% - 106%
    Serum 110 104% - 114%
    Heparin Plasma 109 106% - 111%
    EDTA Plasma 110 105% - 115%
    Citrate Plasma 109 99% - 116%

  • 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

    Abcam’s CD14 in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of CD14 protein in human serum, plasma and cell culture supernatants.


    The 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. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.


     

  • Notes

    CD14 is a LPS-binding glycoprotein primarily expressed on myeloid cells. In response to a gram negative bacterial infection, LPS-activated CD14 leads to the production of inflammatory cytokines. CD14 plays a role in additional cellular functions, including recognition of peptidylglycan from gram positive cells and binding of apoptotic cells to induce phagocytosis.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Microplate (12 x 8 well strips)

Properties

Applications

Our Abpromise guarantee covers the use of ab208983 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

  • 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.

     

  • Standard curve comparison between human CD14 SimpleStep ELISA® kit and traditional ELISA kit from leading competitor. SimpleStep ELISA kit shows a 30-fold increase in sensitivity.

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

  • The concentrations of CD14 were measured in duplicates, interpolated from the CD14 standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 1:625, plasma (citrate) 1:800, plasma (heparin) 1:800 and plasma (EDTA) 1:800. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean CD14 concentration was determined to be 897 ng/mL in serum, 1128 ng/mL in plasma (citrate), 1,195 ng/mL in plasma (heparin) and 1,212 ng/mL in plasma (EDTA).

  • The concentrations of CD14 were measured, interpolated from the CD14 standard curves and corrected for sample dilution. Undiluted samples are as follows: PHA stimulated PBMC 25% and unstimulated PBMC 25%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean CD14 concentration was determined to be 7.91 ng/mL in PHA stimulated PBMC and 6.92 ng/mL in unstimulated PBMC.

  • Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean CD14 concentration was determined to be 1,360 ng/mL with a range of 429 – 2,955 ng/mL.

Protocols

References

ab208983 has not yet been referenced specifically in any publications.

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