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Human GM-CSF DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
Assay Range
15.6 - 1,000 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 human 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)

Data Example

Human GM-CSF ELISA Standard Curve

Product Datasheets

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 Human 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.

41 Citations: Showing 1 - 10
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  1. Repeated exposure of bronchial epithelial cells to particular matter increases allergen-induced cytokine release and permeability
    Authors: H Janbazacya, J van Bergen, S Varasteh, J Garssen, G Folkerts, S Braber
    Cytokine, 2022;154(0):155878.
    Species: Human
    Sample Types: Cell Culture Supernates
  2. CRISPR/Cas9-mediated TGFbetaRII disruption enhances anti-tumor efficacy of human chimeric antigen receptor T cells in vitro
    Authors: K Alishah, M Birtel, E Masoumi, L Jafarzadeh, HR Mirzaee, J Hadjati, RH Voss, M Diken, S Asad
    Journal of Translational Medicine, 2021;19(1):482.
    Species: Human
    Sample Types: Cell Culture Supernates
  3. Gain-of-function variants in SYK cause immune dysregulation and systemic inflammation in humans and mice
    Authors: L Wang, D Aschenbren, Z Zeng, X Cao, D Mayr, M Mehta, M Capitani, N Warner, J Pan, L Wang, Q Li, T Zuo, S Cohen-Keda, J Lu, RC Ardy, DJ Mulder, D Dissanayak, K Peng, Z Huang, X Li, Y Wang, X Wang, S Li, S Bullers, AN Gammage, K Warnatz, AI Schiefer, G Krivan, V Goda, WHA Kahr, M Lemaire, Genomics E, CY Lu, I Siddiqui, MG Surette, D Kotlarz, KR Engelhardt, HR Griffin, R Rottapel, H Decaluwe, RM Laxer, M Proietti, S Hambleton, S Elcombe, CH Guo, B Grimbacher, I Dotan, SC Ng, SA Freeman, SB Snapper, C Klein, K Boztug, Y Huang, D Li, HH Uhlig, AM Muise
    Nature Genetics, 2021;53(4):500-510.
    Species: Human
    Sample Types: Cell Culture Supernates
  4. Altered plasma levels of &betaC and &gammaC chain cytokines and post-treatment modulation in tuberculous lymphadenitis
    Authors: GR Kathamuthu, K Moideen, R Sridhar, D Baskaran, S Babu
    Cytokine, 2021;138(0):155405.
    Species: Human
    Sample Types: Plasma
  5. Characterization of low-density granulocytes in COVID-19
    Authors: LE Cabrera, PT Pekkarinen, M Alander, KHA Nowlan, NA Nguyen, S Jokiranta, S Kuivanen, A Patjas, S Mero, SH Pakkanen, S Heinonen, A Kantele, O Vapalahti, E Kekäläinen, T Strandin
    PloS Pathogens, 2021;17(7):e1009721.
    Species: Human
    Sample Types: Serum
  6. IL-6 promotes drug resistance through formation of polyploid giant cancer cells and stromal fibroblast reprogramming
    Authors: N Niu, J Yao, RC Bast, AK Sood, J Liu
    Oncogenesis, 2021;10(9):65.
    Species: Human
    Sample Types: Cell Culture Supernates
  7. A digital single-molecule nanopillar SERS platform for predicting and monitoring immune toxicities in immunotherapy
    Authors: J Li, A Wuethrich, AAI Sina, HH Cheng, Y Wang, A Behren, PN Mainwaring, M Trau
    Nature Communications, 2021;12(1):1087.
    Species: Human
    Sample Types: Serum
  8. Severe T cell hyporeactivity in ventilated COVID-19 patients correlates with prolonged virus persistence and poor outcomes
    Authors: K Renner, T Schwittay, S Chaabane, J Gottschlin, C Müller, C Tiefenböck, JN Salewski, F Winter, S Buchtler, S Balam, MV Malferthei, M Lubnow, D Lunz, B Graf, F Hitzenbich, F Hanses, H Poeck, M Kreutz, E Orsó, R Burkhardt, T Niedermair, C Brochhause, A Gessner, B Salzberger, M Mack
    Nature Communications, 2021;12(1):3006.
    Species: Human
    Sample Types: Cell Culture Supernates
  9. Biomarkers for Comorbidities Modulate the Activity of T-Cells in COPD
    Authors: K Jamal Jame, WJ Gallert, SD Yanik, S Panek, J Kronsbein, D Jungck, A Koch, J Knobloch
    International Journal of Molecular Sciences, 2021;22(13):.
    Species: Human
    Sample Types: Cell Culture Supernates
  10. Bacterial genotoxins induce T�cell senescence
    Authors: SL Mathiasen, L Gall-Mas, IS Pateras, SDP Theodorou, MRJ Namini, MB Hansen, OCB Martin, CK Vadivel, K Ntostoglou, D Butter, M Givskov, C Geisler, AN Akbar, VG Gorgoulis, T Frisan, N Ødum, T Krejsgaard
    Cell Reports, 2021;35(10):109220.
    Species: Human
    Sample Types: Cell Culture Supernates
  11. Altered plasma levels of &betaC and &gammaC chain cytokines and post-treatment modulation in tuberculous lymphadenitis
    Authors: GR Kathamuthu, K Moideen, R Sridhar, D Baskaran, S Babu
    Cytokine, 2020;138(0):155405.
    Species: Human
    Sample Types: Plasma
  12. LPS Induces GM-CSF Production by Breast Cancer MDA-MB-231 Cells via Long-Chain Acyl-CoA Synthetase 1
    Authors: F Al-Rashed, R Thomas, A Al-Roub, F Al-Mulla, R Ahmad
    Molecules, 2020;25(20):.
    Species: Human
    Sample Types: Cell Culture Supernates
  13. T helper cells in synovial fluid of patients with rheumatoid arthritis primarily have a Th1 and a CXCR3+Th2 phenotype
    Authors: J Aldridge, AH Ekwall, L Mark, B Bergström, K Andersson, I Gjertsson, AC Lundell, A Rudin
    Arthritis Res Ther, 2020;22(1):245.
    Species: Human
    Sample Types: Plasma
  14. A new cytokine-based dynamic stratification during induction is highly predictive of survivals in acute myeloid leukemia
    Authors: P Peterlin, J Gaschet, T Guillaume, A Garnier, M Eveillard, A Le Bourgeo, M Cherel, C Debord, Y Le Bris, O Theisen, C Godon, B Mahé, V Dubruille, S Wuilleme, C Touzeau, T Gastinne, N Blin, A Lok, B Tessoulin, S Le Gouill, P Moreau, MC Béné, P Chevallier
    Cancer Medicine, 2020;0(0):.
    Species: Human
    Sample Types: Serum
  15. Historical BCG vaccination combined with drug treatment enhances inhibition of mycobacterial growth ex vivo in human peripheral blood cells
    Authors: SA Prabowo, A Zelmer, L Stockdale, U Ojha, SG Smith, K Seifert, HA Fletcher
    Sci Rep, 2019;9(1):4842.
    Species: Human
    Sample Types: Cell Culture Supernates
  16. The human glomerular endothelial cells are potent pro-inflammatory contributors in an in vitro model of lupus nephritis
    Authors: P Dimou, RD Wright, KL Budge, A Midgley, SC Satchell, M Peak, MW Beresford
    Sci Rep, 2019;9(1):8348.
    Species: Human
    Sample Types: Cell Culture Supernates
  17. Inhibition of IRE1 RNase activity modulates the tumor cell secretome and enhances response to chemotherapy
    Authors: SE Logue, EP McGrath, P Cleary, S Greene, K Mnich, A Almanza, E Chevet, RM Dwyer, A Oommen, P Legembre, F Godey, EC Madden, B Leuzzi, J Obacz, Q Zeng, JB Patterson, R Jäger, AM Gorman, A Samali
    Nat Commun, 2018;9(1):3267.
    Species: Human
    Sample Types: Cell Culture Supernates
  18. 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;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  19. Caspase-8 Acts in a Non-enzymatic Role as a Scaffold for Assembly of a Pro-inflammatory FADDosome Complex upon TRAIL Stimulation
    Authors: CM Henry, SJ Martin
    Mol. Cell, 2017;65(4):715-729.e5.
    Species: Human
    Sample Types: Cell Culture Supernates
  20. Cross-Tissue Transcriptomic Analysis of Human Secondary Lymphoid Organ-Residing ILC3s Reveals a Quiescent State in the Absence of Inflammation
    Authors: YE Bar-Ephrai, F Cornelisse, N Papazian, T Konijn, RM Hoogenboez, MA Sanders, BA Westerman, M Gönültas, J Kwekkeboom, JMM Den Haan, RM Reijmers, RE Mebius, T Cupedo
    Cell Rep, 2017;21(3):823-833.
    Species: Human
    Sample Types: Cell Culture Supernates
  21. A microfluidics assay to study invasion of human placental trophoblast cells
    Authors: Y Abbas, CM Oefner, WJ Polacheck, L Gardner, L Farrell, A Sharkey, R Kamm, A Moffett, ML Oyen
    J R Soc Interface, 2017;14(130):.
    Species: Human
    Sample Types: Cell Culture Supernates
  22. CREB regulates TNF-?-induced GM-CSF secretion via p38 MAPK in human lung fibroblasts
    Authors: Y Koga, T Hisada, T Ishizuka, M Utsugi, A Ono, M Yatomi, Y Kamide, H Aoki-Saito, H Tsurumaki, K Dobashi, M Yamada
    Allergol Int, 2016;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  23. IL-17 Promotes Neutrophil-Mediated Immunity by Activating Microvascular Pericytes and Not Endothelium
    J Immunol, 2016;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  24. Chronic Endotoxemia in Subjects with Type-1 Diabetes Is Seen Much before the Onset of Microvascular Complications.
    Authors: Aravindhan V, Mohan V, Arunkumar N, Sandhya S, Babu S
    PLoS ONE, 2015;10(9):e0137618.
    Species: Human
    Sample Types: Serum
  25. Roles for the mitogen-activated protein kinase (MAPK) phosphatase, DUSP1, in feedback control of inflammatory gene expression and repression by dexamethasone.
    Authors: Shah S, King E, Chandrasekhar A, Newton R
    J Biol Chem, 2014;289(19):13667-79.
    Species: Human
    Sample Types: Cell Culture Supernates
  26. Activin A as a mediator of NK-dendritic cell functional interactions.
    Authors: Seeger P, Bosisio D, Parolini S, Badolato R, Gismondi A, Santoni A, Sozzani S
    J Immunol, 2014;192(3):1241-8.
    Species: Human
    Sample Types: Cell Culture Supernates
  27. Leptin up-regulates TLR2 in human monocytes.
    Authors: Jaedicke K, Roythorne A, Padget K, Todryk S, Preshaw P, Taylor J
    J Leukoc Biol, 2013;93(4):561-71.
    Species: Human
    Sample Types: Cell Culture Supernates
  28. Opposite cytokine synthesis by fibroblasts in contact co-culture with osteosarcoma cells compared with transwell co-cultures.
    Authors: David M, Kelly E, Zoellner H
    Cytokine, 2013;62(1):48-51.
    Species: Human
    Sample Types: Cell Culture Supernates
  29. ERK5 protein promotes, whereas MEK1 protein differentially regulates, the Toll-like receptor 2 protein-dependent activation of human endothelial cells and monocytes.
    Authors: Wilhelmsen K, Mesa K, Lucero J, Xu F, Hellman J
    J Biol Chem, 2012;287(32):26478-94.
    Species: Human
    Sample Types: Cell Culture Supernates
  30. Interleukin-1alpha controls allergic sensitization to inhaled house dust mite via the epithelial release of GM-CSF and IL-33.
    J. Exp. Med., 2012;209(8):1505-17.
    Species: Human
    Sample Types: Cell Culture Supernates
  31. The role of IL-33 and its receptor ST2 in human nasal epithelium with allergic rhinitis.
    Authors: Kamekura R, Kojima T, Takano K, Go M, Sawada N, Himi T
    Clin. Exp. Allergy, 2012;42(2):218-28.
    Species: Human
    Sample Types: Cell Culture Supernates
  32. Novel markers of inflammation identified in tumor necrosis factor receptor-associated periodic syndrome (TRAPS) by transcriptomic analysis of effects of TRAPS-associated tumor necrosis factor receptor type I mutations in an endothelial cell line.
    Authors: Rebelo SL, Amel-Kashipaz MR, Radford PM, Bainbridge SE, Fiets R, Fang J, McDermott EM, Powell RJ, Todd I, Tighe PJ
    Arthritis Rheum., 2009;60(1):269-80.
    Species: Human
    Sample Types: Cell Culture Supernates
  33. Functional regulatory T cells accumulate in aged hosts and promote chronic infectious disease reactivation.
    Authors: Lages CS, Suffia I, Velilla PA, Huang B, Warshaw G, Hildeman DA, Belkaid Y, Chougnet C
    J. Immunol., 2008;181(3):1835-48.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  34. Fungal proteases induce Th2 polarization through limited dendritic cell maturation and reduced production of IL-12.
    Authors: Lamhamedi-Cherradi SE, Martin RE, Ito T, Kheradmand F, Corry DB, Liu YJ, Moyle M
    J. Immunol., 2008;180(9):6000-9.
    Species: Human
    Sample Types: Cell Culture Supernates
  35. Human CD4(+) T cells recognize an epitope within alpha-fetoprotein sequence and develop into TGF-beta-producing CD4(+) T cells.
    Authors: Alisa A, Boswell S, Pathan AA, Ayaru L, Williams R, Behboudi S
    J. Immunol., 2008;180(7):5109-17.
    Species: Human
    Sample Types: Cell Culture Supernates
  36. Human epithelial cells establish direct antifungal defense through TLR4-mediated signaling.
    Authors: Weindl G, Naglik JR, Kaesler S, Biedermann T, Hube B, Korting HC, Schaller M
    J. Clin. Invest., 2007;117(12):3664-3672.
    Species: Human
    Sample Types: Cell Culture Supernates
  37. Expression and role of EGFR ligands induced in airway cells by PM2.5 and its components.
    Authors: Rumelhard M, Ramgolam K, Hamel R, Marano F, Baeza-Squiban A
    Eur. Respir. J., 2007;30(6):1064-73.
    Species: Human
    Sample Types: Cell Culture Supernates
  38. B and T lymphocytes are the primary sources of RANKL in the bone resorptive lesion of periodontal disease.
    Authors: Kawai T, Matsuyama T, Hosokawa Y, Makihira S, Seki M, Karimbux NY, Goncalves RB, Valverde P, Dibart S, Li YP, Miranda LA, Ernst CW, Izumi Y, Taubman MA
    Am. J. Pathol., 2006;169(3):987-98.
    Species: Human
    Sample Types: Tissue Homogenates
  39. Differential survival of leukocyte subsets mediated by synovial, bone marrow, and skin fibroblasts: site-specific versus activation-dependent survival of T cells and neutrophils.
    Authors: Filer A, Parsonage G, Smith E, Osborne C, Thomas AM, Curnow SJ, Rainger GE, Raza K, Nash GB, Lord J, Salmon M, Buckley CD
    Arthritis Rheum., 2006;54(7):2096-108.
    Species: Human
    Sample Types: Cell Culture Supernates
  40. Activation of resting human primary T cells with chimeric receptors: costimulation from CD28, inducible costimulator, CD134, and CD137 in series with signals from the TCR zeta chain.
    Authors: Finney HM, Akbar AN, Lawson AD
    J. Immunol., 2004;172(1):104-13.
    Species: Human
    Sample Types: Cell Culture Supernates
  41. Adherence influences monocyte responsiveness to interleukin-10.
    Authors: Petit-Bertron AF, Fitting C, Cavaillon JM, Adib-Conquy M
    J. Leukoc. Biol., 2003;73(1):145-54.
    Species: Human
    Sample Types: Cell Culture Supernates

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Reviews for Human GM-CSF DuoSet ELISA

Average Rating: 5 (Based on 5 Reviews)

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Human GM-CSF DuoSet ELISA
By Anonymous on 09/07/2021
Sample Tested: 293T human embryonic kidney cell line

The ELISA worked well. Easy to prepare it.


Human GM-CSF DuoSet ELISA
By Robert Berahovich on 09/29/2020
Sample Tested: Tissue Culture Media

Detection of GM-CSF released by human CAR-T cells producing GM-CSF (PMC655) compared to normal CAR-T cells (PMC650).


Human GM-CSF DuoSet ELISA
By Anonymous on 09/16/2019
Sample Tested: Cell culture supernatant

Human GM-CSF DuoSet ELISA
By Andrew Riley on 09/02/2019
Sample Tested: Tissue Culture Media

human thyroid explant material cultured in a microfluidic device


Human GM-CSF DuoSet ELISA
By Prakasha Kempaiah on 06/03/2016
Sample Tested: Serum