• Assay Type
    Solid Phase Sandwich ELISA
  • Format
    96-well strip plate
  • Assay Length
    3.5 hours or 4.5 hours
  • Sample Type & Volume Required Per Well
    Cell Culture Supernates (100 uL), Serum (100 uL), EDTA Plasma (100 uL), Heparin Plasma (100 uL), Citrate Plasma (100 uL)
  • Sensitivity
    20 pg/mL
  • Assay Range
    39.0 - 2,500 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma)
  • Specificity
    Natural and recombinant human G-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.
Control Available
QC01-1, Quantikine Immunoassay Control Group 1 - Please Inquire
Product Summary
The Quantikine Human G-CSF Immunoassay is a 3.5 or 4.5 hour solid phase ELISA designed to measure G-CSF in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant human G-CSF and antibodies raised against the protein. It has been shown to accurately quantitate recombinant human G-CSF. Results obtained using natural human G-CSF showed linear 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 natural human G-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, Citrate Plasma
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020202020
Mean280827169617610942169
Standard Deviation7.814.119.57.334.681.6
CV%2.81.71.14.13.23.8

Cell Culture Supernates
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020202020
Mean17751510471438181514
Standard Deviation4.59.422177277.1
CV%2.51.82.111.98.85.1

Recovery

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

Sample Type Average % Recovery Range %
Cell Culture Media (n=5) 94 85-110
Citrate Plasma (n=5) 90 78-111
EDTA Plasma (n=5) 93 80-117
Heparin Plasma (n=5) 87 76-103
Serum (n=5) 88 75-106
Linearity
To assess the linearity of the assay, five samples were spiked with high concentrations of G-CSF in various matrices and diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human G-CSF Quantikine ELISA Kit
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: G-CSF
G-CSF (granulocyte-colony stimulating factor) is a secreted glycoprotein that regulates the proliferation, differentiation, and activation of neutrophilic granulocyte lineage cells. It also enhances M-CSF induced monocyte differentiation and the proliferation of Th2-inducing dendritic cells. G-CSF promotes the development of T cell immune tolerance as well as tissue recovery following myocardial infarction and cerebral ischemia. It signals through G-CSF R/CD114 on monocytes, neutrophils, megakaryocytes, platelets, myeloid progenitors, trophoblasts and placenta, endothelial cells, and various tumor cell types. Mutations in G-CSF R are associated with congenital neutropenia.
    • Long Name
      Granulocyte Colony Stimulating Factor
    • Entrez Gene IDs
      1440 (Human); 12985 (Mouse);
    • Alternate Names
      C17orf33; chromosome 17 open reading frame 33; colony stimulating factor 3 (granulocyte); CSF3; CSF3OS; Filgrastim; GCSF; G-CSF; GCSFlenograstim; granulocyte colony-stimulating factor; Lenograstim; MGC45931; pluripoietin;
    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. 100 µL Assay Diluent
    4.   Add 100 µL of Assay Diluent to each well.

    5. 100 µL Standard, Control, or Sample
    6.   Add 100 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 twice for a total of 3 washes.

    8. 200 µL Conjugate
    9.   Add 200 µL of Conjugate to each well.
    10.   For Serum & Plasma Samples: Cover with a new plate sealer, and incubate at room temperature for 2 hours.
      For Cell Culture Supernate Samples: Cover with a new plate sealer, and incubate at room temperature for 1 hour.
    11.   Aspirate and wash 3 times.

    12. 200 µL Substrate Solution
    13.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 20 minutes. PROTECT FROM LIGHT.

    14. 50 µL Stop Solution
    15. Add 50 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
    Filter your results:

    Species
    Sample Type
    1. Profiling of cytokines, chemokines and other soluble proteins as a potential biomarker in colorectal cancer and polyps
      Authors: NA Johdi, L Mazlan, I Sagap, R Jamal
      Cytokine, 2017;99(0):35-42.
      Species: Human
      Sample Type: Serum
    2. Human lung fibroblasts may modulate dendritic cell phenotype and function: results from a pilot in vitro study
      Authors: O Freynet, J Marchal-So, F Jean-Louis, A Mailleux, B Crestani, P Soler, L Michel
      Respir Res, 2016;17(1):36.
      Species: Human
      Sample Type: Cell Culture Supernates
    3. Enhanced non-viral gene delivery by coordinated endosomal release and inhibition of ?-tubulin deactylase
      Nucleic Acids Res, 2016;0(0):.
      Species: Human
      Sample Type: Cell Culture Supernates
    4. Regulation of Circulating Hematopoietic Stem/Progenitor Cells in Preterm Infants with Septicemia
      Stem Cells Dev, 2016;0(0):.
      Species: Human
      Sample Type: Plasma
    5. LPS-Induced G-CSF Expression in Macrophages Is Mediated by ERK2, but Not ERK1.
      Authors: Chang S, Lin S, Yang H, Chou Y, Gao J, Lu S
      PLoS ONE, 2015;10(6):e0129685.
      Species: Human
      Sample Type: Cell Culture Supernates
    6. Interleukin 4, interleukin 6 and osteopontin-serological markers of head and neck malignancy in primary diagnostics: A pilot study.
      Authors: Aderhold C, Grobschmidt G, Sauter A, Faber A, Hormann K, Schultz J
      Oncol Lett, 2014;8(3):1112-1118.
      Species: Human
      Sample Type: Serum
    7. Granulocyte colony-stimulating factor receptor signalling via Janus kinase 2/signal transducer and activator of transcription 3 in ovarian cancer.
      Authors: Kumar, J, Fraser, F W, Riley, C, Ahmed, N, McCulloch, D R, Ward, A C
      Br J Cancer, 2014;110(1):133-45.
      Species: Human
      Sample Type: Cell Culture Supernates
    8. Activation of critical, host-induced, metabolic and stress pathways marks neutrophil entry into cystic fibrosis lungs.
      Authors: Makam M, Diaz D, Laval J, Gernez Y, Conrad CK, Dunn CE, Davies ZA, Moss RB, Herzenberg LA, Herzenberg LA, Tirouvanziam R
      Proc. Natl. Acad. Sci. U.S.A., 2009;106(14):5779-83.
      Species: Human
      Sample Type: Plasma
    9. Age-dependent mobilization of circulating endothelial progenitor cells in infants and young children undergoing cardiac surgery with cardiopulmonary bypass.
      Authors: Sun Y, Yi D, Wang Y, Zheng R, Sun G, Wang J, Liu Y, Ren J, Wang Y, Zhang S, Gu C, Pei J
      Cytokine, 2009;47(3):206-13.
      Species: Human
      Sample Type: Plasma
    10. 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: Human
      Sample Type: Cell Culture Supernates
    11. Hematopoietic progenitor cell mobilization and harvesting in children with malignancies: do the advantages of pegfilgrastim really translate into clinical benefit?
      Authors: Merlin E, Zohar S, Jérôme C, Veyrat-Masson R, Marceau G, Paillard C, Auvrignon A, Le Moine P, Gandemer V, Sapin V, Halle P, Boiret-Dupre N, Chevret S, Demeocq F, Dubray C, Kanold J
      Bone Marrow Transplant., 2009;43(12):919-25.
      Species: Human
      Sample Type: Serum
    12. Randomized trial and pharmacokinetic study of pegfilgrastim versus filgrastim after dose-intensive chemotherapy in young adults and children with sarcomas.
      Authors: Fox E, Widemann BC, Hawkins DS, Jayaprakash N, Dagher R, Aikin AA, Bernstein D, Long L, Mackall C, Helman L, Steinberg SM, Balis FM
      Clin. Cancer Res., 2009;15(23):7361-7.
      Species: Human
      Sample Type: Serum
    13. Does granulocyte colony-stimulating factor ameliorate the proinflammatory response in human meningococcal septic shock?
      Authors: Rojahn A, Brusletto B, Ovstebo R, Haug KB, Kierulf P, Brandtzaeg P
      Crit. Care Med., 2008;36(9):2583-9.
      Species: Human
      Sample Type: Plasma
    14. Staphylococcus aureus in nasal lavage and biopsy of patients with chronic rhinosinusitis.
      Authors: Niederfuhr A, Kirsche H, Deutschle T, Poppert S, Riechelmann H, Wellinghausen N
      Allergy, 2008;63(10):1359-67.
      Species: Human
      Sample Type: Nasal Lavage
    15. Isolation and characterization of CD146+ multipotent mesenchymal stromal cells.
      Authors: Sorrentino A, Ferracin M, Castelli G, Biffoni M, Tomaselli G, Baiocchi M, Fatica A, Negrini M, Peschle C, Valtieri M
      Exp. Hematol., 2008;36(8):1035-46.
      Species: Human
      Sample Type: Cell Culture Supernates
    16. Innate immune responses to TREM-1 activation: overlap, divergence, and positive and negative cross-talk with bacterial lipopolysaccharide.
      Authors: Dower K, Ellis DK, Saraf K, Jelinsky SA, Lin LL
      J. Immunol., 2008;180(5):3520-34.
      Species: Human
      Sample Type: Cell Culture Supernates
    17. Microvascular endothelial cells increase proliferation and inhibit apoptosis of native human acute myelogenous leukemia blasts.
      Authors: Hatfield K, Ryningen A, Corbascio M, Bruserud O
      Int. J. Cancer, 2006;119(10):2313-21.
      Species: Human
      Sample Type: Cell Culture Supernates
    18. Proinflammatory mediators elicit secretion of the intracellular B-lymphocyte stimulator pool (BLyS) that is stored in activated neutrophils: implications for inflammatory diseases.
      Authors: Scapini P, Carletto A, Nardelli B, Calzetti F, Roschke V, Merigo F, Tamassia N, Pieropan S, Biasi D, Sbarbati A, Sozzani S, Bambara L, Cassatella MA
      Blood, 2005;105(2):830-7.
      Species: Human
      Sample Type: Cell Culture Supernates
    19. Is granulocyte colony-stimulating factor level predictive for human IVF outcome?
      Authors: Salmassi A, Schmutzler AG, Schaefer S, Koch K, Hedderich J, Jonat W, Mettler L
      Hum. Reprod., 2005;20(9):2434-40.
      Species: Human
      Sample Type: Follicular Fluid
    20. Enhanced circulating half-life and hematopoietic properties of a human granulocyte colony-stimulating factor/immunoglobulin fusion protein.
      Authors: Cox GN, Smith DJ, Carlson SJ, Bendele AM, Chlipala EA, Doherty DH
      Exp. Hematol., 2004;32(5):441-9.
      Species: Human
      Sample Type: Plasma
    21. Spontaneous circulation of myeloid-lymphoid-initiating cells and SCID-repopulating cells in sickle cell crisis.
      Authors: Lamming CE, Augustin L, Blackstad M, Lund TC, Hebbel RP, Verfaillie CM
      J. Clin. Invest., 2003;111(6):811-9.
      Species: Human
      Sample Type: Serum
    22. Retroviral interleukin 1alpha gene transfer in bone marrow stromal cells in a primate model: induction of myelopoiesis stimulation.
      Authors: de Revel T, Becard N, Sorg T, Rousseau S, Spano JP, Thiebot H, Methali M, Gras G, Le Grand R, Dormont D
      Br. J. Haematol., 2002;118(3):875-84.
      Species: Primate - Macaca fascicularis (Crab-eating Monkey or Cynomolgus Macaque)
      Sample Type: Cell Culture Supernates
    23. Distinct hematopoietic support by two human stromal cell lines.
      Authors: Loeuillet C, Bernard G, Remy-Martin J, Saas P, Herve P, Douay L, Chalmers D
      Exp. Hematol., 2001;29(6):736-45.
      Species: Human
      Sample Type: Cell Culture Supernates
    24. Glioblastoma causing granulocytosis by secretion of granulocyte-colony-stimulating factor.
      Authors: Hintzen RQ, Voormolen J, Sonneveld P, van Duinen SG
      Neurology, 2000;54(1):259-61.
      Species: Human
      Sample Type: CSF
    25. Macrophages that have ingested apoptotic cells in vitro inhibit proinflammatory cytokine production through autocrine/paracrine mechanisms involving TGF-beta, PGE2, and PAF.
      Authors: Fadok, V A, Bratton, D L, Konowal, A, Freed, P W, Westcott, J Y, Henson, P M
      J Clin Invest, 1998;101(4):890-8.
      Species: Human
      Sample Type: Cell Culture Supernates
    Expand to show all 25 Citations
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    Other Experimental Details

    Other Experimental DetailsHuman G-CSF Quantikine ELISA kit is used for Diagnostic use.

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