Overview

  • Product name
    Human IL-8 ELISA Kit
    See all IL-8 kits
  • Detection method
    Colorimetric
  • Precision
    Intra-assay
    Sample n Mean SD CV%
    Overall 5 1.8%
    Inter-assay
    Sample n Mean SD CV%
    Overall 3 7.5%
  • Sample type
    Cell culture supernatant, Serum, Heparin Plasma, EDTA Plasma, Citrate Plasma
  • Assay type
    Sandwich (quantitative)
  • Sensitivity
    1.8 pg/ml
  • Range
    3.91 pg/ml - 250 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

    Human IL8 SimpleStep ELISA kit (ab174442) has been re-developed with new capture and detector antibodies.  This new kit will have the same name but a different product number (ab214030). We have identified new recombinant monoclonal antibodies to use in the SimpleStep ELISA® platform that provide a higher sensitivity when quantification of IL8 in human serum, plasma and cell culture supernatants.


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


    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

    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
    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
    ab221824 - Antibody Diluent 4BI 1 x 6ml
    Human IL-8 Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Sample Diluent NS 1 x 50ml
    Stop Solution 1 x 12ml
    TMB Development Solution 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 chemotactic factor
    • Monocyte-derived neutrophil-activating peptide
    • NAF
    • NAP-1
    • NAP1
    • Neutrophil activating peptide 1
    • Neutrophil activating protein 1
    • Neutrophil-activating factor
    • Neutrophil-activating protein 1
    • Protein 3 10C
    • Protein 3-10C
    • SCYB8
    • Small inducible cytokine subfamily B member 8
    • T cell chemotactic factor
    • T-cell chemotactic factor
    • TSG1
    see all
  • Database links

Associated products

Applications

Our Abpromise guarantee covers the use of ab214030 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
  • Standard curve comparison between human IL-8 SimpleStep ELISA® kit and traditional ELISA kit from leading competitor. SimpleStep ELISA® kit shows a 3-fold increase in sensitivity.

  • 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.

  • 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).

  • 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. Mean of duplicate values +/-SD are graphed. 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

ab214030 has not yet been referenced specifically in any publications.

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