Mouse IL-3 Quantikine ELISA Kit

Catalog # Availability Size / Price Qty
M3000
Mouse IL-3 ELISA Standard Curve
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Product Details
Procedure
Citations (13)
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Mouse IL-3 Quantikine ELISA Kit Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 uL), Serum (50 uL)
Sensitivity
2.5 pg/mL
Assay Range
7.8 - 500 pg/mL (Cell Culture Supernates, Serum)
Specificity
Natural and recombinant mouse IL-3
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
Interference
No significant interference observed with available related molecules.

Product Summary

The Quantikine Mouse IL-3 Immunoassay is a 4.5 hour solid phase ELISA designed to measure mouse IL-3 in cell culture supernates and mouse serum. It contains E. coli-expressed recombinant mouse IL-3 and antibodies raised against recombinant mouse IL-3. This Immunoassay has been shown to accurately quantitate the recombinant factor. Results obtained using natural mouse IL-3 showed dose curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for naturally occurring mouse IL-3.

Precision

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision

Cell Culture Supernates, Serum

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean 33 119 396 32 120 400
Standard Deviation 1.7 8.3 22 2.5 5.5 18
CV% 5.2 7 5.6 7.8 4.6 4.5

Recovery

The recovery of mouse IL-3 spiked to three levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Supernates (n=5) 104 91-114
Serum (n=5) 96 90-106

Linearity

To assess the linearity of the assay, five or more samples containing and/or spiked with various concentrations of mouse IL-3 in each matrix were diluted with Calibrator Diluent and then assayed.
Mouse IL-3 ELISA Linearity

Data Examples

Mouse IL-3 ELISA Standard Curve

Product Datasheets

Preparation and Storage

Shipping
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: IL-3

M-CSF, also known as CSF-1, is a four-alpha-helical-bundle cytokine that is the primary regulator of macrophage survival, proliferation and differentiation. M-CSF is also essential for the survival and proliferation of osteoclast progenitors. M-CSF also primes and enhances macrophage killing of tumor cells and microorganisms, regulates the release of cytokines and other inflammatory modulators from macrophages, and stimulates pinocytosis. M-CSF increases during pregnancy to support implantation and growth of the decidua and placenta. Sources of M-CSF include fibroblasts, activated macrophages, endometrial secretory epithelium, bone marrow stromal cells and activated endothelial cells. The M-CSF receptor (c-fms) transduces its pleotropic effects and mediates its endocytosis. M-CSF mRNAs of various sizes occur. Full length human M-CSF transcripts encode a 522 amino acid (aa) type I transmembrane (TM) protein with a 464 aa extracellular region, a 21 aa TM domain, and a 37 aa cytoplasmic tail that forms a 140 kDa covalent dimer. Differential processing produces two proteolytically cleaved, secreted dimers. One is an N- and O- glycosylated 86 kDa dimer, while the other is modified by both glycosylation and chondroitin-sulfate proteoglycan (PG) to generate a 200 kDa subunit. Although PG-modified M-CSF can circulate, it may be immobilized by attachment to type V collagen. Shorter transcripts encode M-CSF that lack cleavage and PG sites and produce an N-glycosylated 68 kDa TM dimer and a slowly produced 44 kDa secreted dimer. Although forms may vary in activity and half-life, all contain the N-terminal 150 aa portion that is necessary and sufficient for interaction with the M-CSF receptor. The first 223 aa of mature human M-CSF shares 88%, 86%, 81% and 74% aa identity with corresponding regions of dog, cow, mouse and rat M-CSF, respectively. Human M-CSF is active in the mouse, but mouse M-CSF is reported to be species-specific.

Long Name:
Interleukin 3
Entrez Gene IDs:
3562 (Human); 16187 (Mouse); 24495 (Rat)
Alternate Names:
Hematopoietic growth factor; IL3; IL-3; IL-3MGC79398; interleukin 3 (colony-stimulating factor, multiple); interleukin-3; Mast cell growth factor; mast-cell growth factor; MCGF; MCGFMGC79399; MULTI-CSF; multilineage-colony-stimulating factor; Multipotential colony-stimulating factor; P-cell stimulating factor; P-cell-stimulating factor
⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.

Assay Procedure

Refer to the product for complete assay procedure.

Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
  1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
  2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

  3. 50 µL Assay Diluent
  4.   Add 50 µL of Assay Diluent to each well.

  5. 50 µL Standard, Control, or Sample
  6.   Add 50 µL of Standard, Control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
  7.   Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.

  8. 100 µL Conjugate
  9.   Add 100 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
  10.   Aspirate and wash 5 times.

  11. 100 µL Substrate Solution
  12.   Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.

  13. 100 µL Stop Solution
  14.   Add 100 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.

Citations for Mouse IL-3 Quantikine ELISA Kit

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

13 Citations: Showing 1 - 10
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  1. Impact of eosinophil-peroxidase (EPX) deficiency on eosinophil structure and function in mouse airways
    Authors: CM Percopo, JO Krumholz, ER Fischer, LS Kraemer, M Ma, K Laky, HF Rosenberg
    J. Leukoc. Biol., 2018;0(0):.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  2. Calf Spleen Extractive Injection protects mice against cyclophosphamide-induced hematopoietic injury through G-CSF-mediated JAK2/STAT3 signaling
    Authors: W Lu, D Jia, S An, M Mu, X Qiao, Y Liu, X Li, D Wang
    Sci Rep, 2017;7(1):8402.
    Species: Mouse
    Sample Types: Plasma
  3. Comprehensive Ex Vivo Transposon Mutagenesis Identifies Genes That Promote Growth Factor Independence and Leukemogenesis.
    Authors: Guo Y, Updegraff B, Park S, Durakoglugil D, Cruz V, Maddux S, Hwang T, O'Donnell K
    Cancer Res, 2015;76(4):773-86.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  4. Protein malnutrition induces bone marrow mesenchymal stem cells commitment to adipogenic differentiation leading to hematopoietic failure.
    Authors: Cunha M, Lima F, Vinolo M, Hastreiter A, Curi R, Borelli P, Fock R
    PLoS ONE, 2013;8(3):e58872.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  5. Bcl3 prevents acute inflammatory lung injury in mice by restraining emergency granulopoiesis.
    Authors: Kreisel D, Sugimoto S, Tietjens J, Zhu J, Yamamoto S, Krupnick AS, Carmody RJ, Gelman AE
    J. Clin. Invest., 2011;121(1):265-76.
    Species: Mouse
    Sample Types: Serum
  6. Suppression of established experimental autoimmune uveitis by anti-LFA-1alpha Ab.
    Authors: Ke Y, Sun D, Zhang P, Jiang G, Kaplan HJ, Shao H
    Invest. Ophthalmol. Vis. Sci., 2007;48(6):2667-75.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  7. Regulatory T cells prevent catastrophic autoimmunity throughout the lifespan of mice.
    Authors: Kim JM, Rasmussen JP, Rudensky AY
    Nat. Immunol., 2007;8(2):191-7.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  8. IL-7 induces myelopoiesis and erythropoiesis.
    Authors: Aiello FB, Keller JR, Klarmann KD, Dranoff G, Mazzucchelli R, Durum SK
    J. Immunol., 2007;178(3):1553-63.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  9. A novel polysaccharide of black soybean promotes myelopoiesis and reconstitutes bone marrow after 5-flurouracil- and irradiation-induced myelosuppression.
    Authors: Liao HF, Chen YJ, Yang YC
    Life Sci., 2005;77(4):400-13.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  10. Increased interleukin-10 associated with low IL-6 concentration correlated with greater survival rates in mice infected by rabies virus vaccinated against it and immunomodulated with P. acnes.
    Authors: Megid J, Kaneno R, Nozaki CN, Brito CJ, Almeida MF
    Comp. Immunol. Microbiol. Infect. Dis., 2004;27(6):393-411.
    Species: Mouse
    Sample Types: Serum
  11. Interleukin 3 and granulocyte-macrophage colony-stimulating factor are not required for induction of chronic myeloid leukemia-like myeloproliferative disease in mice by BCR/ABL.
    Authors: Li S, Gillessen S, Tomasson MH, Dranoff G, Gilliland DG, Van Etten RA
    Blood, 2001;97(5):1442-50.
    Species: Mouse
    Sample Types: Serum
  12. The protein tyrosine kinase inhibitor AG126 prevents the massive microglial cytokine induction by pneumococcal cell walls.
    Authors: Hanisch UK, Prinz M, Angstwurm K, Hausler KG, Kann O, Kettenmann H, Weber JR
    Eur. J. Immunol., 2001;31(7):2104-15.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  13. A secreted and LIF-mediated stromal cell-derived activity that promotes ex vivo expansion of human hematopoietic stem cells.
    Authors: Shih CC, Hu MC, Hu J
    Blood, 2000;95(6):1957-66.
    Species: Mouse
    Sample Types: Cell Culture Supernates

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