CatchPoint SimpleStep ELISA

Human C Reactive Protein ELISA Kit, Fluorescent (ab229409)

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Overview

  • Product name

    Human C Reactive Protein ELISA Kit, Fluorescent
    See all C Reactive Protein kits

  • Detection method

    Fluorescent

  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Serum 5 1.5%
    Inter-assay
    Sample n Mean SD CV%
    Serum 2 7.2%

  • Sample type

    Cell culture supernatant, Serum, Hep Plasma, EDTA Plasma, Cit plasma

  • Assay type

    Sandwich (quantitative)

  • Sensitivity

    1.9 pg/ml

  • Range

    3.91 pg/ml - 1000 pg/ml

  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Cell culture media 90.1 88.3% - 93.4%

  • Assay time

    1h 30m

  • Assay duration

    One step assay

  • Species reactivity

    Reacts with: Human

  • Product overview

    C-Reactive Protein (CRP) in vitro CatchPoint SimpleStep ELISA (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of C-Reactive Protein (CRP) protein in human serum, plasma, and cell culture supernatants.


    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

    CRP displays several functions associated with host defense: it promotes agglutination, bacterial capsular swelling, phagocytosis and complement fixation through its calcium-dependent binding to phosphorylcholine. CRP can interact with DNA and histones and it may scavenge nuclear material released from damaged circulating cells. CRP is secreted; it forms a homopentamer pentaxin (or pentraxin) which have a discoid arrangement of 5 non-covalently bound subunits. CRP binds 2 calcium ions per subunit. The concentration of CRP in plasma increases greatly during acute phase response to tissue injury, infection or other inflammatory stimuli. It is induced by IL1/interleukin-1 and IL6//interleukin-6.

  • 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 ab229409 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

  • Other - Human C Reactive Protein ELISA Kit, Fluorescent (ab229409)
    Other - Human C Reactive Protein ELISA Kit, Fluorescent (ab229409)

    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 C-Reactive Protein (CRP) standard curve in Sample Diluent NS
    Example of human C-Reactive Protein (CRP) standard curve in Sample Diluent NS

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

  • Titration of pooled human serum within the working range of the assay
    Titration of pooled human serum within the working range of the assay

    Background subtracted data values (mean +/- SD, n=2) are graphed.

  • CRP concentrations in 10 individual human serum donors
    CRP concentrations in 10 individual human serum donors

    10,000X diluted sera from 10 apparently healthy male donors were measured using this kit. Interpolated data values corrected for sample dilution are graphed in ng of CRP per mL of serum (mean +/- SD, n=2). Nine out of ten individual human sera tested within or at reported Human serum range (< 5,000 ng/mL, dotted line). The mean of CRP concentration of these nine individual sera was determined to be 2,240 ng/mL with a range of 559 – 5,285 ng/mL. Note that one individual human serum sample (donor # 2) tested substantially higher, 11,737 ng/mL.

  • Comparison of CRP signals in three healthy and three rheumatoid arthritis patient sera
    Comparison of CRP signals in three healthy and three rheumatoid arthritis patient sera

    Background subtracted data values of two serum dilutions (as indicated in parenthesis) are graphed (mean +/- SD, n=2).

  • Quantification of CRP concentrations in three healthy and three rheumatoid arthritis (RA) patient sera
    Quantification of CRP concentrations in three healthy and three rheumatoid arthritis (RA) patient sera

    The concentrations of CRP were interpolated from data values shown in Figure 4 using CRP standard curve, corrected for sample dilution, and graphed in ng of CRP per mL of serum. As expected, note that the CRP serum concentrations are increased in RA patients.

Protocols

References

ab229409 has not yet been referenced specifically in any publications.

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