Human GM-CSF Quantikine HS ELISA Kit

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HSGM0
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Human GM-CSF ELISA Urine Assay
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Human GM-CSF Quantikine HS ELISA Kit Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
6.5 hours
Sample Type & Volume Required Per Well
Serum (150 uL), EDTA Plasma (150 uL), Heparin Plasma (150 uL), Urine (200 uL)
Sensitivity
0.26 pg/mL
Assay Range
1.0 - 64 pg/mL (Serum, EDTA Plasma, Heparin Plasma), 0.5 - 32 pg/mL (Urine)
Specificity
Natural and recombinant human 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 HS Human GM-CSF Immunoassay is a 6.5 hour solid phase ELISA designed to measure human GM-CSF in serum, plasma, and urine. It contains E. coli-expressed recombinant human GM-CSF and antibodies raised against the recombinant factor. It has been shown to accurately quantitate the recombinant factor. Results obtained using natural GM-CSF showed linear curves that were parallel to the standard curves obtained using the Quantikine HS kit standards. These results indicate that this kit can be used to determine relative mass values for natural human 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

Serum, EDTA Plasma, Heparin Plasma

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 18 18 18
Mean (pg/mL) 3.68 22 44.6 2.8 16.2 33.2
Standard Deviation 0.35 1.07 1.76 0.66 2.47 4.6
CV% 9.5 4.9 4 23.5 15.3 13.9

Urine

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (pg/mL) 2.69 7.05 24.8 3.77 14.3 27.4
Standard Deviation 0.14 0.33 0.73 0.37 1.17 1.85
CV% 5.2 4.7 2.9 9.8 8.2 6.8

Recovery

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

Sample Type Average % Recovery Range %
EDTA Plasma (n=4) 114 88-140
Heparin Plasma (n=4) 109 91-122
Serum (n=4) 109 94-128
Urine (n=5) 102 95-116

Linearity

To assess the linearity of the assay, samples were spiked with high concentrations of GM-CSF in various matrices and diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human GM-CSF High Sensitivity ELISA Linearity

Data Examples

Human GM-CSF ELISA Urine Assay

Human GM-CSF ELISA Serum/Plasma Assay

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.
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

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. Assay Diluent
  4.   For Serum & Plasma Samples: Add 100 µL of Assay Diluent to each well.
    For Urine Samples: Add 50 µL of Assay Diluent to each well.

  5. Standard, Control, or Sample
  6.   For Serum & Plasma Samples: Add 150 µL of Standard, control, or sample to each well.
    For Urine Samples: Add 200 µL of Standard, control, or sample to each well.
  7.   Cover with a plate sealer, and incubate at room temperature for 3 hours.
  8.   Aspirate each well and wash, repeating the process 5 times for a total of 6 washes.

  9. 200 µL Conjugate
  10.   Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
  11.   Aspirate and wash 6 times.

  12. 50 µL Substrate Solution
  13.   Add 50 µL Substrate Solution to each well. Cover with a new plate sealer, and incubate at room temperature for 1 hour. Do not wash the plate.

  14. 50 µL Amplifier Solution
  15. Add 50 µL Amplifier Solution to each well. Cover with a new plate sealer, and incubate at room temperature for 30 minutes.

  16. 50 µL Stop Solution
  17. Add 50 µL of Stop Solution to each well. Read at 490 nm within 30 minutes. Set wavelength correction to 650 nm or 690 nm.

Citations for Human GM-CSF Quantikine HS 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.

11 Citations: Showing 1 - 10
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  1. Effects of Ingestion of Different Amounts of Carbohydrate after Endurance Exercise on Circulating Cytokines and Markers of Neutrophil Activation
    Authors: K Tanisawa, K Suzuki, S Ma, S Kondo, S Okugawa, M Higuchi
    Antioxidants (Basel), 2018;7(4):.
    Species: Human
    Sample Types: Plasma
  2. Decreased Count and Dysfunction of Circulating EPCs in Postmenopausal Hypercholesterolemic Females via Reducing NO Production
    Authors: Y Luo, QN Yan, WZ Wu, FY Luo
    Stem Cells Int, 2018;2018(0):2543847.
    Species: Human
    Sample Types: Plasma
  3. Immunogenicity of a novel Clade B HIV-1 vaccine combination: Results of phase 1 randomized placebo controlled trial of an HIV-1 GM-CSF-expressing DNA prime with a modified vaccinia Ankara vaccine boost in healthy HIV-1 uninfected adults
    Authors: SP Buchbinder, NA Grunenberg, BJ Sanchez, KE Seaton, G Ferrari, MA Moody, N Frahm, DC Montefiori, CM Hay, PA Goepfert, LR Baden, HL Robinson, X Yu, PB Gilbert, MJ McElrath, Y Huang, GD Tomaras,
    PLoS ONE, 2017;12(7):e0179597.
    Species: Human
    Sample Types: Serum
  4. 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 Types: Plasma
  5. Safety and immunologic correlates of Melanoma GVAX, a GM-CSF secreting allogeneic melanoma cell vaccine administered in the adjuvant setting.
    Authors: Lipson E, Sharfman W, Chen S, McMiller T, Pritchard T, Salas J, Sartorius-Mergenthaler S, Freed I, Ravi S, Wang H, Luber B, Sproul J, Taube J, Pardoll D, Topalian S
    J Transl Med, 2015;13(0):214.
    Species: Human
    Sample Types: Serum
  6. A randomized trial of recombinant human granulocyte-macrophage colony stimulating factor for patients with acute lung injury.
    Authors: Paine R, Standiford TJ, Dechert RE, Moss M, Martin GS, Rosenberg AL, Thannickal VJ, Burnham EL, Brown MB, Hyzy RC
    Crit. Care Med., 2012;40(1):90-7.
    Species: Human
    Sample Types: BALF
  7. Timed sequential treatment with cyclophosphamide, doxorubicin, and an allogeneic granulocyte-macrophage colony-stimulating factor-secreting breast tumor vaccine: a chemotherapy dose-ranging factorial study of safety and immune activation.
    Authors: Emens LA, Asquith JM, Leatherman JM, Kobrin BJ, Petrik S, Laiko M, Levi J, Daphtary MM, Biedrzycki B, Wolff AC, Stearns V, Disis ML, Ye X, Piantadosi S, Fetting JH, Davidson NE, Jaffee EM
    J. Clin. Oncol., 2009;27(35):5911-8.
    Species: Human
    Sample Types: Serum
  8. Duffy (Fy), DARC, and neutropenia among women from the United States, Europe and the Caribbean.
    Authors: Grann VR, Ziv E, Joseph CK, Neugut AI, Wei Y, Jacobson JS, Horwitz MS, Bowman N, Beckmann K, Hershman DL
    Br. J. Haematol., 2008;143(2):288-93.
    Species: Human
    Sample Types: Plasma
  9. Neutropenia in 6 ethnic groups from the Caribbean and the U.S.
    Authors: Grann VR, Bowman N, Joseph C, Wei Y, Horwitz MS, Jacobson JS, Santella RP, Hershman DL
    Cancer, 2008;113(4):854-60.
    Species: Human
    Sample Types: Plasma
  10. Increased circulating endothelial progenitor cells in septic patients: correlation with survival.
    Authors: Rafat N, Hanusch C, Brinkkoetter PT, Schulte J, Brade J, Zijlstra JG, van der Woude FJ, van Ackern K, Yard BA, Beck GCh
    Crit. Care Med., 2007;35(7):1677-84.
    Species: Human
    Sample Types: Serum
  11. Endotoxin-induced maturation of monocytes in preterm fetal sheep lung.
    Authors: Kramer BW, Joshi SN, Moss TJ, Newnham JP, Sindelar R, Jobe AH, Kallapur SG
    Am. J. Physiol. Lung Cell Mol. Physiol., 2007;293(2):L345-53.
    Species: Ovine
    Sample Types: BALF

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