Mouse GM-CSF Quantikine ELISA Kit

(25 citations)   
  • 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
    5.8 pg/mL
  • Assay Range
    7.8 - 500 pg/mL (Cell Culture Supernates, Serum)
  • Specificity
    Natural and recombinant mouse GM-CSF
  • 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 GM-CSF Immunoassay is a 4.5 hour solid phase ELISA designed to measure mouse GM-CSF in cell culture supernates and serum. It contains E. coli-expressed recombinant mouse GM-CSF and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate the recombinant mouse GM-CSF accurately. Results obtained using natural mouse GM-CSF showed dose-response curves that were parallel to the standard curves obtained using the recombinant standards. These results indicate that this kit can be used to determine relative mass values for natural mouse GM-CSF.

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, Plasma
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020202020
Mean18.567.22192071.7243
Standard Deviation0.72.16.913.811.2
CV%3.83.13.255.34.6

Recovery

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

Sample Type Average % Recovery Range %
Cell Culture Supernates (n=8) 96 82-114
Serum (n=7) 94 82-112
Linearity
To assess the linearity of the assay, five or more samples containing and/or spiked with various concentrations of mouse GM-CSF in each matrix were diluted with Calibrator Diluent and then assayed.
Mouse GM-CSF Quantikine ELISA Kit
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: GM-CSF
GM-CSF (Granulocyte-Macrophage Colony Stimulating Factor) induces the development of monocytes, neutrophils, eosinophils, and myeloid and dermal dendritic cells. It also acts as a neutrophil and dendritic cell chemoattractant. GM-CSF promotes Th1 and Th17 cell mediated autoimmune inflammation as well as the inflammatory activation of dendritic cells, microglia, alveolar macrophages, and eosinophils. In addition, it cooperates with G-CSF in promoting tumor cell proliferation and invasion. GM-CSF signals through a receptor complex composed of GM-CSF R alpha and the Common beta Chain (beta c) with Syndecan-2 as a potential co-receptor. The beta c subunit is shared by the receptor complexes for IL-3 and IL-5.
    • Long Name
      Granulocyte Macrophage Growth Factor
    • Entrez Gene IDs
      1437 (Human); 12981 (Mouse); 116630 (Rat); 397208 (Porcine); 403923 (Canine); 493805 (Feline);
    • Alternate Names
      colony stimulating factor 2 (granulocyte-macrophage); Colony-stimulating factor; CSF; CSF2; GMCSF; GM-CSF; GMCSFgranulocyte-macrophage colony-stimulating factor; granulocyte-macrophage colony stimulating factor; MGC131935; MGC138897; molgramostin; sargramostim;
    Related Research Areas
    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:

    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.

    25 Citations: Showing 1 - 10
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    Species
    Sample Type
    1. Adipose progenitor cell secretion of GM-CSF and MMP9 promotes a stromal and immunological microenvironment that supports breast cancer progression
      Authors: F Reggiani, V Labanca, P Mancuso, C Rabascio, G Talarico, S Orecchioni, A Manconi, F Bertolini
      Cancer Res., 2017;0(0):.
      Species: Mouse
      Sample Type: 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 Type: Plasma
    3. A heart-brain-kidney network controls adaptation to cardiac stress through tissue macrophage activation
      Authors: K Fujiu, M Shibata, Y Nakayama, F Ogata, S Matsumoto, K Noshita, S Iwami, S Nakae, I Komuro, R Nagai, I Manabe
      Nat. Med., 2017;23(5):611-622.
      Species: Mouse
      Sample Type: Serum
    4. Modifying Antigen-Encapsulating Liposomes with KALA Facilitates MHC Class I Antigen Presentation and Enhances Anti-tumor Effects
      Authors: N Miura, H Akita, N Tateshita, T Nakamura, H Harashima
      Mol. Ther, 2017;0(0):.
      Species: Mouse
      Sample Type: Serum
    5. Development and Preclinical Application of an Immunocompetent Transplant Model of Basal Breast Cancer with Lung, Liver and Brain Metastases
      PLoS ONE, 2016;11(5):e0155262.
      Species: Mouse
      Sample Type:
    6. Key role of microRNA in the regulation of granulocyte macrophage colony-stimulating factor expression in murine alveolar epithelial cells during oxidative stress.
      Authors: Sturrock, Anne, Mir-Kasimov, Mustafa, Baker, Jessica, Rowley, Jesse, Paine, Robert 3
      J Biol Chem, 2014;289(7):4095-105.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    7. Oncolytic viral therapy with a combination of HF10, a herpes simplex virus type 1 variant and granulocyte-macrophage colony-stimulating factor for murine ovarian cancer.
      Authors: Goshima F, Esaki S, Luo C, Kamakura M, Kimura H, Nishiyama Y
      Int J Cancer, 2014;134(12):2865-77.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    8. 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 Type: Cell Culture Supernates
    9. 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 Type: Serum
    10. Keratinocyte Growth Factor Augments Pulmonary Innate Immunity through Epithelium-driven, GM-CSF-dependent Paracrine Activation of Alveolar Macrophages.
      Authors: Wu H, Suzuki T, Carey B, Trapnell BC, McCormack FX
      J. Biol. Chem., 2011;286(17):14932-40.
      Species: Mouse
      Sample Type: Tissue Homogenates
    11. Deletion of tristetraprolin caused spontaneous reactive granulopoiesis by a non-cell-autonomous mechanism without disturbing long-term hematopoietic stem cell quiescence.
      Authors: Kaplan IM, Morisot S, Heiser D, Cheng WC, Kim MJ, Civin CI
      J. Immunol., 2011;186(5):2826-34.
      Species: Mouse
      Sample Type: Plasma
    12. Histone Deacetylase Inhibitor LAQ824 Augments Inflammatory Responses in Macrophages through Transcriptional Regulation of IL-10.
      Authors: Wang H, Cheng F
      J. Immunol., 2011;186(0):3986.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    13. Macrophage tumor necrosis factor-alpha induces epithelial expression of granulocyte-macrophage colony-stimulating factor: impact on alveolar epithelial repair.
      Authors: Cakarova L, Marsh LM, Wilhelm J, Mayer K, Grimminger F, Seeger W, Lohmeyer J, Herold S
      Am. J. Respir. Crit. Care Med., 2009;180(6):521-32.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    14. Novel immunocompetent murine tumor model for evaluation of conditionally replication-competent (oncolytic) murine adenoviral vectors.
      Authors: Robinson M, Li B, Ge Y, Ko D, Yendluri S, Harding T, VanRoey M, Spindler KR, Jooss K
      J. Virol., 2009;83(8):3450-62.
      Species: Mouse
      Sample Type: Tissue Homogenates
    15. Simultaneous absence of surfactant proteins A and D increases lung inflammation and injury after allogeneic HSCT in mice.
      Authors: Gram K, Yang S, Steiner M, Somani A, Hawgood S, Blazar BR, Panoskaltsis-Mortari A, Haddad IY
      Am. J. Physiol. Lung Cell Mol. Physiol., 2009;296(2):L167-75.
      Species: Mouse
      Sample Type: BALF
    16. 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 Type: Cell Culture Supernates
    17. Administration of cyclooxygenase-2 inhibitor reduces joint inflammation but exacerbates osteopenia in IL-1 alpha transgenic mice due to GM-CSF overproduction.
      Authors: Niki Y, Takaishi H, Takito J, Miyamoto T, Kosaki N, Matsumoto H, Toyama Y, Tada N
      J. Immunol., 2007;179(1):639-46.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    18. Lentiviral vector-mediated autonomous differentiation of mouse bone marrow cells into immunologically potent dendritic cell vaccines.
      Authors: Koya RC, Kimura T, Ribas A, Rozengurt N, Lawson GW, Faure-Kumar E, Wang HJ, Herschman H, Kasahara N, Stripecke R
      Mol. Ther., 2007;15(5):971-80.
      Species: Mouse
      Sample Type: Serum
    19. GM-CSF and the impaired pulmonary innate immune response following hyperoxic stress.
      Authors: Baleeiro CE, Christensen PJ, Morris SB, Mendez MP, Wilcoxen SE, Paine R
      Am. J. Physiol. Lung Cell Mol. Physiol., 2006;291(6):L1246-55.
      Species: Mouse
      Sample Type: Tissue Homogenates
    20. Expression of CCL20 and granulocyte-macrophage colony-stimulating factor, but not Flt3-L, from modified vaccinia virus ankara enhances antiviral cellular and humoral immune responses.
      Authors: Chavan R, Marfatia KA, An IC, Garber DA, Feinberg MB
      J. Virol., 2006;80(15):7676-87.
    21. Organ-specific role of MyD88 for gene regulation during polymicrobial peritonitis.
      Authors: Weighardt H, Mages J, Jusek G, Kaiser-Moore S, Lang R, Holzmann B
      Infect. Immun., 2006;74(6):3618-32.
      Species: Mouse
      Sample Type: Tissue Homogenates
    22. 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 Type: Cell Culture Supernates
    23. Endogenous granulocyte-macrophage colony-stimulating factor overexpression in vivo results in the long-term recruitment of a distinct dendritic cell population with enhanced immunostimulatory function.
      Authors: Miller G, Pillarisetty VG, Shah AB, Lahrs S, Xing Z, DeMatteo RP
      J. Immunol., 2002;169(6):2875-85.
      Species: Mouse
      Sample Type: Serum
    24. 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 Type: Serum
    25. 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 Type: Cell Culture Supernates
    Expand to show all 25 Citations
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