Human IL-3 ELISA Kit
The Human IL-3 ELISA Kit is For Research Use Only
Size: 1×96 wells
Sensitivity: 15 pg/mL
Dynamic Range: 31.25 – 1000 pg/ml
Incubation Time: 3.5 hours
Sample Type: Serum, Plasma, Cell Culture
Sample Size: 100 µl
Alternative Names: Interleukin 3, Mast Cell Growth Factor, P-cell stimulating factor, Burst Promoting Activity, Multicolony Stimulating Factor, t h y-1 inducing factor, WEHI-3 growth factor, MULTI-CSF, MCGF, MGC79398, MGC79399
SENSITIVITY, SPECIFICITY AND REPEATABILITY
REPEATABILITY: The coefficient of variation of both intra-assay and inter-assay were less than 10%.
SENSITIVITY: The minimum detectable dose was 15 pg/mL.
SPECIFICITY: This assay recognizes both natural and recombinant human IL-3. The factors listed below were prepared at 10ng/ml in Standard /sample Diluent and assayed for cross-reactivity and no significant cross-reactivity or interference was observed.
Interleukin-3, also known as mast cell growth factor, P-cell stimulating factor, burst promoting activity, multicolony stimulating factor, t h y-1 inducing factor and WEHI-3 growth factor, is a pleiotropic factor produced primarily by activated T cells that can stimulate the proliferation and differentiation of pluripotent hematopoietic stem cells as well as various lineage committed progenitors. In addition, IL-3 also affects the functional activity of mature mast cells, basophils, eosinophils and macrophages. In addition to activated T cells, other cell types such as human thymic epithelial cells, activated murine mast cells, murine keratinocytes and neurons/astrocytes can also produce IL-3. Mature human IL-3 share only 29% aa sequence identity with murine IL-3. IL-3 activity is highly species specific and human IL-3 does not show activity on murine cells. IL-3 is involved in a variety of cell activities such as cell growth, differentiation and apoptosis. This cytokine has been shown to also possess neurotrophic activity, and it may be associated with neurologic disorders.
Human IL-2 ELISA Assay
Human IL-4 ELISA Assay
Human IL-5 ELISA Assay
The Eagle Biosciences Human Interleukin 3 (IL-3) ELISA Assay Kit employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for IL-3 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any IL-3 present is bound by the immobilized antibody. Following incubation unbound samples are removed during a wash step, and then a detection antibody specific for IL-3 is added to the wells and binds to the combination of capture antibody- IL-3 in sample. Following a wash to remove any unbound combination, and enzyme conjugate is added to the wells. Following incubation and wash steps a substrate is added. A colored product is formed in proportion to the amount of IL-3 present in the sample. The reaction is terminated by addition of acid and absorbance is measured at 450nm. A standard curve is prepared from seven IL-3 standard dilutions and IL-3 sample concentration determined.
- Prepare all reagents and working standards as directed in the previous sections.
- Determine the number of microwell strips required to test the desired number of samples plus appropriate number of wells needed for running blanks and standards. Remove extra microwell strips from holder and store in foil bag with the desiccant provided at 2-8°C sealed tightly.
- Add 100 µL of Standard, control, or sample, per well. Cover with the adhesive strip provided. Incubate for 1.5 hours at 37°C
- Aspirate each well and wash, repeating the process three times for a total of four washes. Wash by filling each well with Wash Buffer (350 µL) using a squirt bottle, manifold dispenser or auto-washer. Complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating or decanting. Invert the plate and blot it against clean paper towels.
- Add 100 µL of the working solution of Biotin-Conjugate to each well. Cover with a new adhesive strip and incubate 1 hour at 37°C
- Repeat the aspiration/wash.
- Add 100 µL of the working solution of Streptavidin-HRP to each well. Cover with a new adhesive strip and incubate for 30 minutes at 37°C Avoid placing the plate in direct light.
- Repeat the aspiration/wash.
- Add 100 µL of Substrate Solution to each well. Incubate for 10-20 minutes at 37°C. Avoid placing the plate in direct light.
- Add 100 µL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
- Determine the optical density of each well immediately, using a microplate reader set to 450 nm.(optionally 630nm as the reference wave length;610-650nm is acceptable)
Please note: All documents above are for reference use only and should not be used in place of the documents included with this physical product. If digital copies are needed, please contact us.
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