Overview

  • Product name

    Human KGF ELISA Kit
    See all KGF kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Cell media 8 3.2%
    Inter-assay
    Sample n Mean SD CV%
    Cell media 3 2.5%
  • Sample type

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

    Sandwich (quantitative)
  • Sensitivity

    7.2 pg/ml
  • Range

    31.25 pg/ml - 2000 pg/ml
  • Recovery

    103.54 %

    Sample specific recovery
    Sample type Average % Range
    Serum 103.5 93.5% - 109.8%
    Cell culture media 114.5 105% - 117.3%
    Heparin Plasma 108.4 105.2% - 115.2%
    EDTA Plasma 104.6 86.1% - 109.6%
    Citrate Plasma 110.6 94.5% - 114.5%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Human
  • Product overview

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


    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

    Human Keratinocyte Growth Factor (KGF) is a 28 kDa glycoprotein and the seventh discovered member of the FGF family (FGF-7). It is primarily secreted by cells of mesenchymal origin and it acts in a paracrine fashion on cells expressing the tyrosine kinase receptor FGFR2b. Binding to the receptor occurs in the presence of heparin sulphate glycosaminoglycans leading to the formation of KGF-FGFR2b-HS dimmers which allow trans-phosphorylation of specific cytoplasmic kinases.

    Signaling through FGFR2b has a dual effect, whereby it phosphorylates proteins that promote growth stimulation (FRS2-alpha, IRS4, IGF-1, ERK2, SHP2), as well as proteins that attenuate cell growth and promote cell differentiation (Lamina-associated polypeptide 2, SAP102 and DIg family of tumor suppressor proteins). Furthermore, KGF also induces anti-apoptotic pathways through AKT and p21-activated protein kinase 4 (PAK4) and up-regulates multiple genes involved in proliferation, migration, angiogenesis, differentiation, survival, nucleotide biosynthesis, tissue remodeling, cell cycle progression, DNA repair and detoxification of reactive oxygen species. All these molecular processes lead to tissue protection and regeneration.

     

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Microplate

Properties

Associated products

Applications

Our Abpromise guarantee covers the use of ab183362 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.

     

  • Figure 1. Example of KGF standard curve in sample diluent 25BS. The KGF standard curve was prepared as described in Section 10. Background-subtracted data values (mean +/- SD) are graphed.

  • Figure 2. Specificity of KGF signal on stimulated and non stimulated media supernatants. Human Dermal Neonatal Fibroblasts cells were cultured in HGDMEM supplemented with 10% fetal calf serum, 2 mM L-glutamine, 100 U/mL penicillin and 100 µg/mL streptomycin. Cells were cultured for 2 days at 37°C in the presence or absence of 1 µg/mL of FGF-basic with or without 10 ng/mL of TNF-alpha. The concentrations of KGF were measured after loading neat media on the plate and were then interpolated from a KGF calibration curve on sample diluent 25BS. The mean concentration of KGF (FGF-7) was determined to be undetectable in non stimulated media, 413 pg/mL in FGF-basic stimulated media and 1,016 pg/mL in FGF-basic+TNF-alpha stimulated media.

Protocols

References

ab183362 has not yet been referenced specifically in any publications.

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