Overview

  • Product name

    Human IL-8 ELISA Kit, Fluorescent
    See all IL-8 kits
  • Detection method

    Fluorescent
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Supernatant 5 1.8%
    Inter-assay
    Sample n Mean SD CV%
    Supernatant 3 7.5%
  • Sample type

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

    Sandwich (quantitative)
  • Sensitivity

    0.45 pg/ml
  • Range

    0.49 pg/ml - 2000 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Serum 99 93% - 103%
    Cell culture media 101 98% - 104%
    Heparin Plasma 87 82% - 91%
    EDTA Plasma 88 84% - 92%
    Citrate Plasma 86 83% - 90%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Human
    Does not react with: Mouse, Rat, Cow
  • Product overview

    IL-8 in vitro CatchPoint SimpleStep ELISA (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of IL-8 protein in human serum, plasma, and cell culture supernatant.


    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

    IL-8 is a chemotactic factor that attracts neutrophils, basophils, and T cells, but not monocytes. It is also involved in neutrophil activation. It is released from several cell types in response to an inflammatory stimulus. IL-8(6-77) has a 5-10-fold higher activity on neutrophil activation, IL-8(5-77) has increased activity on neutrophil activation and IL-8(7-77) has a higher affinity to receptors CXCR1 and CXCR2 as compared to IL-8(1-77), respectively

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Pre-coated microplate (12 x 8 well strips)

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    100X Stoplight Red Substrate 1 x 120µl
    10X Human IL-8 Capture Antibody 1 x 600µl
    10X Human IL-8 Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    500X Hydrogen Peroxide (H2O2, 3%) 1 x 50µl
    Antibody Diluent 4BI 1 x 6ml
    Human IL-8 Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NS (ab193972) 1 x 50ml
    SimpleStep Pre-Coated Black 96-Well Microplate 1 unit
    Stoplight Red Substrate Buffer 1 x 12ml
  • Research areas

  • Function

    IL-8 is a chemotactic factor that attracts neutrophils, basophils, and T-cells, but not monocytes. It is also involved in neutrophil activation. It is released from several cell types in response to an inflammatory stimulus. IL-8(6-77) has a 5-10-fold higher activity on neutrophil activation, IL-8(5-77) has increased activity on neutrophil activation and IL-8(7-77) has a higher affinity to receptors CXCR1 and CXCR2 as compared to IL-8(1-77), respectively.
  • Sequence similarities

    Belongs to the intercrine alpha (chemokine CxC) family.
  • Post-translational
    modifications

    Several N-terminal processed forms are produced by proteolytic cleavage after secretion from at least peripheral blood monocytes, leukcocytes and endothelial cells. In general, IL-8(1-77) is referred to as interleukin-8. IL-8(6-77) is the most promiment form.
  • Cellular localization

    Secreted.
  • Information by UniProt
  • Alternative names

    • (Ala-IL-8)77
    • (Ser-IL-8)72
    • 9E3
    • Beta thromboglobulin like protein
    • C-X-C motif chemokine 8
    • CEF-4
    • chemokine, CXC motif, ligand 8
    • CXCL8
    • Emoctakin
    • GCP-1
    • GCP/IL-8 protein I
    • GCP/IL-8 protein II
    • GCP/IL-8 protein III
    • GCP/IL-8 protein IV
    • GCP/IL-8 protein V
    • GCP/IL-8 protein VI
    • GCP1
    • Granulocyte chemotactic protein 1
    • IL-8
    • IL-8(1-77)
    • IL-8(9-77)
    • IL8
    • IL8/NAP1 form I
    • IL8/NAP1 form II
    • IL8/NAP1 form III
    • IL8/NAP1 form IV
    • IL8/NAP1 form V
    • IL8/NAP1 form VI
    • IL8_HUMAN
    • Inteleukin 8
    • LECT
    • LUCT
    • Lymphocyte-derived neutrophil-activating factor
    • LYNAP
    • MDNCF
    • MDNCF-b
    • MDNCF-c
    • MONAP
    • Monocyte derived neutrophil activating peptide
    • Monocyte derived neutrophil chemotactic factor
    • Monocyte-derived neutrophil chemotactic factor
    • Monocyte-derived neutrophil-activating peptide
    • NAF
    • NAP 1
    • NAP-1
    • NAP1
    • Neutrophil activating peptide 1
    • Neutrophil activating protein 1
    • Neutrophil-activating factor
    • Neutrophil-activating protein 1
    • Protein 3 10C
    • Protein 3-10C
    • SCYB 8
    • SCYB8
    • Small inducible cytokine subfamily B member 8
    • T cell chemotactic factor
    • T-cell chemotactic factor
    • TSG 1
    • TSG1
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab229402 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.

     

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

  • The concentrations of IL-8 were measured in duplicates, interpolated from the IL-8 standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean IL-8 concentration was determined to be 63.2 pg/mL in serum.

  • The concentrations of IL-8 were measured in duplicates, interpolated from the IL-8 standard curves and corrected for sample dilution. Undiluted samples are as follows: plasma (EDTA) 50%, plasma (citrate) 50%, and plasma (heparin) 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).

  • Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean IL-8 concentration was determined to be 61.6 pg/mL with a range of 62 – 174.9 pg/mL; two individuals (Donor #3 and Donor #9) measured below the minimal detectable dose.

  • The concentrations of IL-8 were measured in duplicates, interpolated from the IL-8 standard curves and corrected for sample dilution. Undiluted samples are as follows: unstimulated 1:400 and stimulated 1:400. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). Measured values were interpolated from the IL-8 Standard Curve diluted in Sample Diluent NS and corrected for dilution factor. The mean IL-8 concentration was determined to be 2.2 ng/mL in unstimulated, 119.7ng/mL in stimulated, and undetectable in media.

Protocols

References

ab229402 has not yet been referenced specifically in any publications.

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