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Mouse GM-CSF DuoSet ELISA

Catalog #: DY415
26 Citations Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
DY415
DY415-05
Ancillary Products Available
DY008B: DuoSet ELISA Ancillary Reagent Kit 2
DY999B: Substrate Reagent Pack
DY999: Substrate Reagent Pack
Mouse GM-CSF ELISA Standard Curve
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Product Details
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Citations (26)
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Summary Product Features Kit Content Other Reagents Required Data Examples Product Datasheets Preparation and Storage Background

Mouse GM-CSF DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
Assay Range
7.8 - 500 pg/mL
Sufficient Materials
For five or fifteen 96-well plates*
Specificity
Please see the product datasheet

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant mouse GM-CSF. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.

Product Features

  • Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
  • Development protocols are provided to guide further assay optimization
  • Assay can be customized to your specific needs
  • Economical alternative to complete kits

Kit Content

  • Capture Antibody
  • Detection Antibody
  • Recombinant Standard
  • Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)

Other Reagents Required

DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.

The components listed above may be purchased separately:

PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # WA126), or 0.05% Tween® 20 in PBS, pH 7.2-7.4

Reagent Diluent: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered

Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)

Stop Solution: 2 N H2SO4 (Catalog # DY994)

Microplates: R&D Systems (Catalog # DY990)

Plate Sealers: ELISA Plate Sealers (Catalog # DY992)

Scientific Data

N/A Mouse GM-CSF ELISA Standard Curve View Larger

Mouse GM-CSF ELISA Standard Curve

Product Datasheets

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Preparation and Storage

Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & 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; CSF-2; GMCSF; GM-CSF; GMCSFgranulocyte-macrophage colony-stimulating factor; granulocyte-macrophage colony stimulating factor; MGC131935; MGC138897; Molgramostim; molgramostin; Sargramostim

Assay Procedure

GENERAL ELISA PROTOCOL

Plate Preparation

  1. Dilute the Capture Antibody to the working concentration in PBS without carrier protein. Immediately coat a 96-well microplate with 100 μL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
  2. Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 μL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
  3. Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
  4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

Assay Procedure

  1. Add 100 μL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
  2. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  3. Add 100 μL of the Detection Antibody, diluted in Reagent Diluent, to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
  4. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  5. Add 100 μL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  6. Repeat the aspiration/wash as in step 2.
  7. Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  8. Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  9. Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.

Citations for Mouse GM-CSF DuoSet ELISA

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.

26 Citations: Showing 1 - 10
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  1. GM-CSF-mediated epithelial-immune cell crosstalk orchestrates pulmonary immunity to Aspergillus fumigatus
    Authors: Mills, KAM;Westermann, F;Espinosa, V;Rosiek, E;Desai, JV;Aufiero, MA;Guo, Y;Mitchell, KA;Tuzlak, S;De Feo, D;Lionakis, MS;Rivera, A;Becher, B;Hohl, TM;
    bioRxiv : the preprint server for biology
    Species: Mouse
    Sample Types: Tissue Homogenates
  2. IL-18BP mediates the balance between protective and pathological immune responses to Toxoplasma gondii
    Authors: JT Clark, OE Weizman, DL Aldridge, LA Shallberg, J Eberhard, Z Lanzar, D Wasche, JD Huck, T Zhou, AM Ring, CA Hunter
    Cell Reports, 2023-02-23;42(3):112147.
    Species: Mouse
    Sample Types: Serum
  3. Guidelines for mouse and human DC generation
    Authors: MB Lutz, S Ali, C Audiger, SE Autenrieth, L Berod, V Bigley, L Cyran, M Dalod, J Dörrie, D Dudziak, G Flórez-Gra, L Giusiano, GJ Godoy, M Heuer, AB Krug, CHK Lehmann, CT Mayer, SH Naik, S Scheu, G Schreibelt, E Segura, K Seré, T Sparwasser, J Tel, H Xu, M Zenke
    European Journal of Immunology, 2022-10-27;0(0):.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  4. Heterocellular OSM-OSMR signalling reprograms fibroblasts to promote pancreatic cancer growth and metastasis
    Authors: BY Lee, EKJ Hogg, CR Below, A Kononov, A Blanco-Gom, F Heider, J Xu, C Hutton, X Zhang, T Scheidt, K Beattie, A Lamarca, M McNamara, JW Valle, C Jørgensen
    Nature Communications, 2021-12-17;12(1):7336.
    Species: Mouse
    Sample Types: Tissue Homogenates
  5. Inflammatory macrophages exploit unconventional pro-phagocytic integrins for phagocytosis and anti-tumor immunity
    Authors: Z Tang, D Davidson, R Li, MC Zhong, J Qian, J Chen, A Veillette
    Cell Reports, 2021-12-14;37(11):110111.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  6. Gas6/TAM Signalling Negatively Regulates Inflammatory Induction of GM-CSF in Mouse Brain Microglia
    Authors: SE Gilchrist, GM Pennelli, S Hafizi
    Cells, 2021-11-24;10(12):.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  7. Generation and utilization of a HEK-293T murine GM-CSF expressing cell line
    Authors: EK Robinson, S Covarrubia, S Zhou, S Carpenter
    PLoS ONE, 2021-04-09;16(4):e0249117.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  8. GM-CSF Calibrates Macrophage Defense and Wound Healing Programs during Intestinal Infection and Inflammation
    Authors: T Castro-Dop, A Fleming, TW Dennison, JR Ferdinand, K Harcourt, BJ Stewart, Z Cader, ZK Tuong, C Jing, LSC Lok, RJ Mathews, A Portet, A Kaser, S Clare, MR Clatworthy
    Cell Rep, 2020-07-07;32(1):107857.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  9. DNA threads released by activated CD4+ T lymphocytes provide autocrine costimulation
    Authors: M Costanza, PL Poliani, P Portararo, B Cappetti, S Musio, F Pagani, L Steinman, MP Colombo, R Pedotti, S Sangaletti
    Proc. Natl. Acad. Sci. U.S.A., 2019-04-15;0(0):.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  10. Neutrophil GM-CSF receptor dynamics in acute lung injury
    Authors: S De Alessan, GJ Ferguson, AJ Dodd, JK Juss, A Devaprasad, S Piper, O Wyatt, H Killick, DJ Corkill, ES Cohen, A Pandit, TRDJ Radstake, R Simmonds, AM Condliffe, MA Sleeman, AS Cowburn, DK Finch, ER Chilvers
    J. Leukoc. Biol., 2019-04-03;0(0):.
    Species: Mouse
    Sample Types: BALF
  11. Interferon-gamma inhibits seminal plasma induction of colony stimulating factor 2 in mouse and human reproductive tract epithelial cells
    Authors: DJ Sharkey, DJ Glynn, JE Schjenken, KP Tremellen, SA Robertson
    Biol. Reprod., 2018-09-01;0(0):.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  12. Thymic-Specific Serine Protease Limits Central Tolerance and Exacerbates Experimental Autoimmune Encephalomyelitis
    Authors: L Serre, M Girard, A Ramadan, P Menut, N Rouquié, LE Lucca, K Mahiddine, B Leobon, LT Mars, S Guerder
    J. Immunol., 2017-10-23;0(0):.
    Species: Mouse
    Sample Types: Cell Lysates
  13. Treatment with anti-Fc?RI? antibody exacerbates EAE and T-cell immunity against myelin
    Authors: S Musio, M Costanza, PL Poliani, E Fontana, M Cominelli, G Abolafio, L Steinman, R Pedotti
    Neurol Neuroimmunol Neuroinflamm, 2017-04-14;4(3):e342.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  14. Heat shock protein 27-derived atheroprotection involves reverse cholesterol transport that is dependent on GM-CSF to maintain ABCA1 and ABCG1 expression in ApoE(-/-) mice
    Authors: VK Pulakazhi, A Adijiang, T Seibert, YX Chen, C Shi, Z Batulan, ER O'Brien
    FASEB J, 2017-02-23;0(0):.
    Species: Mouse
    Sample Types: Serum
  15. Exon skipping of Fc?RI? eliminates expression of the high-affinity IgE receptor in mast cells with therapeutic potential for allergy
    Proc. Natl. Acad. Sci. U.S.A., 2016-11-21;113(49):14115-14120.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  16. ER? in CD4+ T Cells Is Crucial for Ligand-Mediated Suppression of Central Nervous System Autoimmunity
    J Immunol, 2016-05-13;0(0):.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  17. Induction of autoimmune disease by deletion of CTLA-4 in mice in adulthood
    Authors: K Klocke, S Sakaguchi, R Holmdahl, K Wing
    Proc Natl Acad Sci USA, 2016-04-11;0(0):.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  18. Thrombospondin-1 restrains neutrophil granule serine protease function and regulates the innate immune response during Klebsiella pneumoniae infection.
    Authors: Zhao Y, Olonisakin T, Xiong Z, Hulver M, Sayeed S, Yu M, Gregory A, Kochman E, Chen B, Mallampalli R, Sun M, Silverstein R, Stolz D, Shapiro S, Ray A, Ray P, Lee J
    Mucosal Immunol, 2014-12-10;8(4):896-905.
    Species: Mouse
    Sample Types: Tissue Homogenates
  19. Injectable, spontaneously assembling, inorganic scaffolds modulate immune cells in vivo and increase vaccine efficacy.
    Authors: Kim J, Li W, Choi Y, Lewin S, Verbeke C, Dranoff G, Mooney D
    Nat Biotechnol, 2014-12-08;33(1):64-72.
    Species: Mouse
    Sample Types: Complex Sample Type
  20. Macrophage Dectin-1 Expression Is Controlled by Leukotriene B4 via a GM-CSF/PU.1 Axis.
    Authors: Serezani CH, Kane S, Collins L
    J. Immunol., 2012-06-13;189(2):906-15.
    Species: Mouse
    Sample Types: BALF
  21. The role of alveolar epithelial cells in initiating and shaping pulmonary immune responses: communication between innate and adaptive immune systems.
    Authors: Chuquimia OD, Petursdottir DH, Rahman MJ, Hartl K, Singh M, Fernandez C
    PLoS ONE, 2012-02-29;7(2):e32125.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  22. IL-33-responsive lineage- CD25+ CD44(hi) lymphoid cells mediate innate type 2 immunity and allergic inflammation in the lungs.
    Authors: Bartemes KR, Iijima K, Kobayashi T, Kephart GM, McKenzie AN, Kita H
    J. Immunol., 2011-12-23;188(3):1503-13.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  23. Local delivery of GM-CSF protects mice from lethal pneumococcal pneumonia.
    Authors: Steinwede K, Tempelhof O, Bolte K, Maus R, Bohling J, Ueberberg B, Langer F, Christman JW, Paton JC, Ask K, Maharaj S, Kolb M, Gauldie J, Welte T, Maus UA
    J. Immunol., 2011-10-14;187(10):5346-56.
    Species: Mouse
    Sample Types: BALF
  24. The encephalitogenicity of T(H)17 cells is dependent on IL-1- and IL-23-induced production of the cytokine GM-CSF.
    Authors: El-Behi M, Ciric B, Dai H
    Nat. Immunol., 2011-04-24;12(6):568-75.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  25. The p110delta catalytic isoform of PI3K is a key player in NK-cell development and cytokine secretion.
    Authors: Kim N, Saudemont A, Webb L, Camps M, Ruckle T, Hirsch E, Turner M, Colucci F
    Blood, 2007-07-20;110(9):3202-8.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  26. A synthetic analog of alpha-galactosylceramide induces macrophage activation via the TLR4-signaling pathways.
    Authors: Hung LC, Lin CC, Hung SK, Wu BC, Jan MD, Liou SH, Fu SL
    Biochem. Pharmacol., 2007-03-12;73(12):1957-70.
    Species: Mouse
    Sample Types: Cell Culture Supernates

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