Overview

  • Product name

    Human IL-3 ELISA Kit
    See all IL-3 kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Serum 5 5.7%
    Inter-assay
    Sample n Mean SD CV%
    Serum 3 4.9%
  • Sample type

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

    Sandwich (quantitative)
  • Sensitivity

    6.7 pg/ml
  • Range

    15 pg/ml - 1000 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Cell culture supernatant 102 99% - 105%
    Urine 91 88% - 92%
    Serum 88 84% - 91%
    Cell culture media 99 94% - 103%
    Heparin Plasma 86 81% - 92%
    EDTA Plasma 89 87% - 89%
    Citrate Plasma 85 84% - 87%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Human
  • Product overview

    IL-3 in vitro SimpleStep ELISA (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of human IL-3 protein in serum, plasma, urine, 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

    Interleukin 3 (IL-3) is a secreted 133 amino acids long, glycosylated interleukin coded by the IL-3 gene. Previous names reported for IL-3 are Hematopoietic growth factor, Mast cell growth factor, multipotenial colony-stimulating factor, and P-cell-stimulating factor. IL-3 has been found to stimulate a variety of cell activities such as; proliferation, differentiation, survival of pluripotent hematopoietic stem cells, and may be associated with neurologic disorders. IL-3 is expressed by activated T-cells, mast cells, and natural killer cells. IL-3 performs its biological action by binding to a transmembrane receptor complex consisting of IL-3RA, also known as CD123, and a signal inducing beta subunit, which also acts as a receptor for colony stimulating factor 2 and Interleukin 5.

  • 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 Wash Buffer PT (ab206977) 1 x 20ml
    Antibody Diluent CPI 1 x 6ml
    10X Human IL-3 Capture Antibody 1 x 600µl
    10X Human IL-3 Detector Antibody 1 x 600µl
    Human IL-3 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

    Granulocyte/macrophage colony-stimulating factors are cytokines that act in hematopoiesis by controlling the production, differentiation, and function of 2 related white cell populations of the blood, the granulocytes and the monocytes-macrophages.
    This CSF induces granulocytes, macrophages, mast cells, stem cells, erythroid cells, eosinophils and megakaryocytes.
  • Tissue specificity

    Activated T-cells, mast cells, natural killer cells.
  • Sequence similarities

    Belongs to the IL-3 family.
  • Cellular localization

    Secreted.
  • Information by UniProt
  • Alternative names

    • Colony stimulating factor multiple
    • Hematopoietic growth factor
    • IL 3
    • IL-3
    • IL3
    • IL3_HUMAN
    • Interleukin 3
    • Interleukin 3 (colony stimulating factor, multiple)
    • Interleukin-3
    • Mast cell growth factor
    • MCGF
    • MGC79398
    • MGC79399
    • Multi CSF
    • Multilineage colony stimulating factor
    • Multipotential colony stimulating factor
    • Multipotential colony-stimulating factor
    • OTTHUMP00000065963
    • P cell stimulating factor
    • P-cell-stimulating factor
    see all
  • Database links

Associated products

Applications

Our Abpromise guarantee covers the use of ab215088 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 IL-3 SimpleStep ELISA® kit and traditional ELISA kit from leading competitor. SimpleStep ELISA kit shows comparable sensitivity with shorter protocol time.

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

  • Human PBMCs were cultured in RPMI supplemented with 10% fetal calf serum, 2mM 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 PHAM. The concentrations of IL-3 were measured in duplicates, interpolated from the IL-3 standard curves and corrected for sample dilution. Undiluted samples are as follows: stimulated supernatant 100% and unstimulated supernatant 100%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean (IL-3 concentration was determined to be 662 pg/mL in 100% stimulated supernatant, and 9.5 pg/mL in 100% unstimulated supernatant.

  • The concentrations of IL-3 were measured in duplicate and interpolated from the IL-3 standard curve and corrected for sample dilution. Undiluted samples are as follows: serum 100%, plasma (citrate) 50%, plasma (EDTA) 50%, plasma (Heparin) 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).

  • The concentrations of IL-3 were measured in duplicate and interpolated from the IL-3 standard curve and corrected for sample dilution. Undiluted samples are as follows: RPMI+10% FBS 50%, urine 100%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).

     

Protocols

References

ab215088 has not yet been referenced specifically in any publications.

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