Overview

  • Product name

    Human CXCL1 ELISA Kit
    See all GRO alpha kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Supernatant 3 1.9%
    Inter-assay
    Sample n Mean SD CV%
    Supernatant 8 3.7%
  • Sample type

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

    Sandwich (quantitative)
  • Sensitivity

    0.064 pg/ml
  • Range

    2.34 pg/ml - 150 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Cell culture supernatant 95 92% - 97%
    Serum 83 82% - 85%
    Heparin Plasma 86 85% - 87%
    Citrate Plasma 81 80% - 82%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

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

    As of November 2, 2018, Human CXCL1 SimpleStep ELISA® kit has been re-developed. We have identified new recombinant monoclonal antibodies to provide improved performance and consistency when quantifying CXCL1 protein in human serum, platelet poor 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 Development Solution 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.


    CXCL1 (GRO alpha) is a member of the CXC family of chemokines. CXCL1 contains the ELR motif and preferentially chemoattracts neutrophils. Human CXCL1 undergoes post-translational modification into three isoforms called CXCL1(4-73), CXCL1(5-73) and CXCL1(6-73). These isoforms are produced by proteolytic cleavage after secretion from peripheral blood monocytes, and all show higher activity in-vitro than the full-length protein. Two additional CXC proteins, CXCL2 (or GRO beta) and CXCL3 (or GRO gamma), share 90% and 86% amino acid sequence homology, respectively, with CXCL1. Human CXCL1 mRNA is expressed in foreskin fibroblasts, synovial fibroblasts, chondrocytes and osteocytes. Additionally, CXCL1 mRNA has been detected in mammary fibroblasts and epithelial cells, endothelial cells, activated monocytes, macrophages, and neutrophils. CXCL1 is a growth-regulated gene and is overexpressed constitutively in tumorigenic cells. As a result, elevated levels of CXCL1 can be seen in several types of tumors and cancers, including lung cancer and melanoma.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Microplate

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Human CXCL1 Capture Antibody 1 x 600µl
    10X Human CXCL1 Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    Antibody Diluent 5BI 1 x 6ml
    Human CXCL1 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

  • Relevance

    The GRO gene was originally identified by subtractive hybridization studies between normal and tumorigenic Chinese hamster embryo fibroblasts. The hamster cDNA was cloned and used as a probe for cloning of the human GRO cDNA. The GROalpha gene initially cloned from T24 cells and the gene in melanoma cells encoding melanoma growth stimulating protein (MGSA) are identical. Human cells contain three closely related, but distinct GRO genes: GRO alpha, GRO beta, and GRO gamma. GRO beta and GRO gamma share 93% and 82% identity, respectively, with GRO alpha at the nucleotide level. GROs are members of the chemokine alpha family that is characterized by the separation with one amino acid of the first two cysteine residues, C-X-C, in the amino acid sequence. The GRO gene has been mapped to chromosome 4q21. In normal cells, human mRNA GRO expression is found in foreskin fibroblasts, synovial fibroblasts, chondrocytes and osteocytes. Additionally, GRO mRNA has been detected in mammary fibroblasts, mammary epithelial cells, endothelial cells, activated monocytes, macrophages, and neutrophils. Characterization of the GROalpha receptor indicates the presence of low and high affinity receptors on human neutrophils.
  • Cellular localization

    Secreted
  • Alternative names

    • C-X-C motif chemokine 1
    • chemokine (C-X-C motif) ligand 1 (melanoma growth stimulating activity, alpha)
    • fibroblast secretory protein
    • FSP
    • GRO
    • GRO protein, alpha
    • GRO-alpha(1-73)
    • GRO1
    • GRO1 oncogene
    • GRO1 oncogene (melanoma growth stimulating activity, alpha)
    • GRO1 oncogene (melanoma growth-stimulating activity)
    • GROa
    • growth-regulated alpha protein
    • Keratinocyte-derived chemokine, mouse, homolog of
    • Melanoma growth stimulatory activity
    • melanoma growth stimulatory activity alpha
    • MGSA
    • MGSA a
    • MGSA alpha
    • NAP 3
    • Neutrophil-activating protein 3
    • SCYB1
    • Small inducible cytokine subfamily B, member 1
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab190805 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 CXCL1 SimpleStep ELISA kit and traditional ELISA kit from leading competitor. SimpleStep ELISA kit shows increased sensitivity.

  • Standard Curve comparison between new human CXCL1 SimpleStep ELISA kit and original ELISA kit. The current SimpleStep ELISA kit shows increased sensitivity.

  • The CXCL1 standard curve was prepared as described in Section 10. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.

     

  • The concentrations of CXCL1 were measured in duplicates, interpolated from the CXCL1 standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 100%, platelet poor plasma (citrate) 100%, and platelet poor plasma (heparin) 100%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean CXCL1 concentration was determined to be 20.65 pg/mL in serum, 11.10 pg/mL in platelet poor plasma (citrate), and 29.39 pg/mL in platelet poor plasma (Heparin).

     

  • The concentrations of CXCL1 were measured in duplicate and interpolated from the CXCL1 standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean CXCL1 concentration was determined to be 17.53 ng/mL in treated PBMC supernatant.

  • Human PBMCs were cultured in RPMI supplemented with 10% fetal calf serum, 2 mM L-glutamine. Cells were cultured for 2 days at 37˚C in the presence or absence of 1.5% PHA-M. The concentrations of CXCL1 were interpolated from the standard curve in Sample Diluent NS and corrected for sample dilution. The mean CXCL1 concentration was determined to be 5.11 pg/mL in unstimulated PBMC supernatants, and 17533.21 pg/mL in stimulated PBMC supernatants.

     

Protocols

References

This product has been referenced in:

  • Bardi GT  et al. Detection of Inflammation-Related Melanoma Small Extracellular Vesicle (sEV) mRNA Content Using Primary Melanocyte sEVs as a Reference. Int J Mol Sci 20:N/A (2019). Read more (PubMed: 30870978) »
  • Pu Y  et al. Adiponectin promotes human jaw bone marrow mesenchymal stem cell chemotaxis via CXCL1 and CXCL8. J Cell Mol Med 21:1411-1419 (2017). Read more (PubMed: 28176455) »
See all 4 Publications for this product

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