Overview

  • Product name

    Human CXCL5 ELISA Kit
    See all CXCL5 kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Serum 8 3.2%
    Inter-assay
    Sample n Mean SD CV%
    Serum 3 5%
  • Sample type

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

    Sandwich (quantitative)
  • Sensitivity

    2 pg/ml
  • Range

    15.63 pg/ml - 1000 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Serum 89 85% - 96%
    Cell culture media 89 82% - 93%
    Heparin Plasma 105 100% - 107%
    EDTA Plasma 98 94% - 104%
    Citrate Plasma 104 94% - 115%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

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

    CXCL5 in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of human CXCL5 protein in serum, plasma, and cell culture supernatant samples.


    The SimpleStep ELISA® employs an affinity tag labelled 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

    Human CXCL5 is a member of the CXC subfamily of chemokines. Full-length CXCL5 is 114 amino acids (aa) long including a 36 aa signal peptide. This signal peptide is cleaved to produce the 78 aa residue secreted protein (ENA-78). ENA-78 is 52%, 48%, and 51% identical in aa sequence to human GROα, GROβ, and GROγ, respectively. CXCL5 is expressed by fibroblasts and is induced by bacterial lipopolysaccharides. Additionally, this chemokine is a potent chemotaxin involved in neutrophil activation. Furthermore, CXCL5 is produced concomitantly with Interleukin 8 (IL8) in response to stimulation with either Interleukin 1 (IL1) or Tumor Necrosis Factor alpha.

  • 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 CXCL5 Capture Antibody 1 x 600µl
    10X Human CXCL5 Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    Antibody Diluent 5BI 1 x 6ml
    Human CXCL5 Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NS (ab193972) 1 x 50ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Development Solution 1 x 12ml
  • Research areas

  • Function

    Involved in neutrophil activation. In vitro, ENA-78(8-78) and ENA-78(9-78) show a threefold higher chemotactic activity for neutrophil granulocytes.
  • Sequence similarities

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

    N-terminal processed forms ENA-78(8-78) and ENA-78(9-78) are produced by proteolytic cleavage after secretion from peripheral blood monocytes.
  • Cellular localization

    Secreted.
  • Information by UniProt
  • Alternative names

    • AMCFII
    • C-X-C motif chemokine 5
    • C-X-C motif chemokine ligand 5
    • Chemokine (C X C motif) ligand 5
    • chemokine (C-X-C motif) ligand 5
    • Cxcl5
    • CXCL5_HUMAN
    • ENA 78
    • ENA-78 (8-78)
    • ENA-78(1-78)
    • ENA-78(9-78)
    • ENA78
    • Epithelial derived neutrophil activating protein 78
    • Epithelial-derived neutrophil-activating protein 78
    • Lipopolysaccharide-induced CXC chemokine
    • Neutrophil activating peptide ENA 78
    • Neutrophil activating protein 78
    • Neutrophil-activating peptide ENA-78
    • neutrophil-activating protein 78
    • SCYB5
    • Small inducible cytokine B5
    • small inducible cytokine subfamily B (Cys-X-Cys), member 5 (epithelial-derived neutrophil-activating peptide 78)
    • small inducible cytokine subfamily B, member 5
    • Small-inducible cytokine B5
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab212163 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 CSCL5 SimpleStep ELISA® kit and traditional ELISA kit from leading competitor. SimpleStep ELISA kit shows a 6-fold increase in sensitivity.

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

  • The concentrations of CXCL5 were measured in duplicates, interpolated from the CXCL5 standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 100%, plasma (citrate) 50%, plasma (EDTA) 50% and plasma (heparin) 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean CXCL5 concentration was determined to be 622.1 pg/mL in serum, 1,977.3 pg/mL in plasma (citrate), 511.4 pg/mL in plasma (EDTA) and 706.2 pg/mL in plasma (heparin).

  • The concentrations of CXCL5 were measured in duplicates, interpolated from the CXCL5 standard curves and corrected for sample dilution. Undiluted samples are as follows: PBMC media (Donor 1) 10% and PBMC media (Donor 2) 2.5%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean CXCL5 concentration was determined to be 2,702.1 pg/mL in PBMC media (Donor 1), and 21,336.6 pg/mL in PBMC media (Donor 2).

  • Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean CXCL5 concentration was determined to be 641.9 pg/mL with a range of 14.1 – 1,315.6 pg/mL.

  • Half of the volume of each sample was then additionally centrifuged at 10,000 x g for 10 minutes at 4oC to remove platelets (platelet poor). All plasma samples were diluted 1:2 in Sample Diluent NS and measured in duplicate. Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean CXCL5 concentration was determined to be 825 pg/mL in plasma (citrate), 852.0 pg/mL in platelet poor plasma (citrate), 255 pg/mL in plasma (EDTA), 254.2 pg/mL in platelet poor plasma (EDTA), 1,067.8 pg/mL in plasma (heparin), and 1,065.2 pg/mL in platelet poor plasma (heparin).

  • All samples were diluted 1:40 and the concentrations of CXCL5 were measured in duplicates, interpolated from the CXCL5 standard curves and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean CXCL5 concentration was determined to be 1,188.3 pg/mL in PBMC unstimulated media (Donor 1), 2,702.2 pg/mL in PBMC stimulated media (Donor 1), 567.2 pg/mL in PBMC unstimulated media (Donor 2), and 21,325.6 pg/mL in PBMC stimulated media (Donor 2). PBMC samples were cultured in RPMI media with 10% fetal bovine serum and 1% PenStrep (unstimulated samples). Then PBMC samples were stimulated with 1.5% PHA-M for 48 hours.

Protocols

References

ab212163 has not yet been referenced specifically in any publications.

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