Overview

  • Product name

    Mouse GDF-15 ELISA Kit
    See all GDF15 kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    EDTA plasma 8 5%
    Inter-assay
    Sample n Mean SD CV%
    EDTA plasma 3 5.7%
  • Sample type

    Cell culture supernatant, Serum, Hep Plasma, EDTA Plasma, Cit plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    22.2 pg/ml
  • Range

    62.5 pg/ml - 4000 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Serum 76 70% - 82%
    Cell culture media 89 84% - 93%
    Hep Plasma 83 77% - 91%
    EDTA Plasma 82 81% - 84%
    Cit plasma 85 83% - 89%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Mouse
  • Product overview

    GDF-15 in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of mouse GDF-15 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

    The cytokine growth differentiation factor 15 (GDF-15) is one of 15 growth/differentiation factors (GDF-1 to GDF-15), all of which are members of the BMP family of TGF-beta superfamily proteins. These growth/differentiation factors are produced as inactive preproproteins which are then cleaved and assembled into active secreted homodimers. Specifically, mouse GDF-15 is synthesized as a 273 amino acid (aa) proprecursor that contains a cleavable 158 aa glycosylated prodomain (aa 31-188) and a 115 aa mature region (aa 189-303). Over aa sequence 189-303, mouse GDF-15 shares 97% and 68% aa identity with rat and human GDF-15, respectively.

    In mice, GDF-15 is expressed in a diverse population of activated cell types, including hepatocytes, macrophages, Schwann cells, cardiomyocytes, osteoblasts, adipocytes. It is also expressed in the epithelium of the small intestine and mammary gland. GDF proteins have a critical role during embryonic development, particularly in the skeletal, nervous, and muscular systems.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Pre-coated microplate (12 x 8 well strips)

Properties

Applications

Our Abpromise guarantee covers the use of ab216947 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 GDF-15 were measured in duplicates, interpolated from the GDF-15 standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 50%, plasma (citrate) 50%, plasma (EDTA) 50%, and plasma (heparin) 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean GDF-15 concentration was determined to be 4,030 pg/mL in neat serum, 4,261 pg/mL in neat plasma (citrate), 4,385 pg/mL in neat plasma (EDTA), and 3,981 pg/mL in neat plasma (heparin).

  • The concentrations of GDF-15 were measured in duplicate, interpolated from the GDF-15 standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 50%, plasma (citrate) 50%, plasma (EDTA) 50%, and plasma (heparin) 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean GDF-15 concentration was determined to be 837.9 pg/mL in neat serum, 895.2 pg/mL in neat plasma (citrate), 1,079.6 pg/mL in neat plasma (EDTA), and 884.6 pg/mL in neat plasma (heparin).

  • The concentrations of GDF-15 were measured in duplicate, interpolated from the GDF-15 standard curves and corrected for sample dilution. Undiluted samples are as follows: pregnant mouse serum 50%, L929 culture media 100%, and lung culture media 100%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean GDF-15 concentration was determined to be 3,268 pg/mL in neat pregnant mouse serum, 2,223 pg/mL in L929 culture media, and 3,665 pg/mL in lung culture media.

  • Samples were loaded neat (100%). The concentrations of GDF-15 were measured in duplicates and interpolated from the GDF-15 standard curves. The interpolated values are plotted (mean +/- SD, n=2). The mean GDF-15 concentration was determined to be below the limit of detection in unstimulated L929 culture media, 2,246 pg/mL in stimulated L929 culture media, 193 pg/mL in unstimulated J774A.1 culture media, and 583 pg/mL in stimulated J774A.1 culture media. L929 cells were grown for 72 hours in MEM media containing 10% fetal horse serum and 100 µg/mL Kanamycin (unstimulated) plus 1.5% PHA and 10 ng/mL PMA (stimulated). J774A.1 cells were grown for 72 hours in HGDMEM media containing 10% fetal bovine serum and 100 µg/mL Kanamycin (unstimulated) plus 1.5% PHA and 10 ng/mL PMA (stimulated).

Protocols

References

ab216947 has not yet been referenced specifically in any publications.

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