Human Flt-3 Ligand Quantikine ELISA Kit

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  • Assay Type
    Solid Phase Sandwich ELISA
  • Format
    96-well strip plate
  • Assay Length
    4 hours or 4.5 hours
  • Sample Type & Volume Required Per Well
    Cell Culture Supernates (50 uL), Serum (50 uL), EDTA Plasma (50 uL), Heparin Plasma (50 uL), Citrate Plasma (50 uL)
  • Sensitivity
    7 pg/mL
  • Assay Range
    15.6 - 1,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma)
  • Specificity
    Natural and recombinant human Flt-3 Ligand
  • 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
Product Summary
The Quantikine Human Flt-3/Flk-2 Ligand Immunoassay is a 4.0 or 4.5 hour solid phase ELISA designed to measure soluble human Flt-3 Ligand in cell culture supernates, serum, and plasma. It contains NS0-expressed recombinant human Flt-3 Ligand and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate recombinant Flt-3 Ligand accurately. Results obtained measuring natural human Flt-3 Ligand showed dose-response curves that were parallel to the standard curves obtained using the recombinant Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for natural human Flt-3 Ligand.

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
Intra-Assay Precision Inter-Assay Precision
Standard Deviation1.

Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma
Intra-Assay Precision Inter-Assay Precision
Standard Deviation1.


The recovery of Flt-3 Ligand 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) 102 92-118
Citrate Plasma (n=5) 101 94-110
EDTA Plasma (n=5) 101 90-119
Heparin Plasma (n=5) 100 88-112
Serum (n=5) 100 91-117
To assess the linearity of the assay, five samples were spiked with high concentrations of Flt-3 Ligand in various matrices and diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human Flt-3 Ligand Quantikine ELISA Kit
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: Flt-3 Ligand
Flt-3 Ligand, also known as FL, is an alpha-helical cytokine that promotes the differentiation of multiple hematopoietic cell lineages. Mature human Flt-3 Ligand consists of a 158 amino acid (aa) extracellular domain (ECD) with a cytokine-like domain and a juxtamembrane tether region, a 21 aa transmembrane segment, and a 30 aa cytoplasmic tail. Within the ECD, human Flt-3 Ligand shares 71% and 65% aa sequence identity with mouse and rat Flt-3 Ligand, respectively. Human and mouse Flt-3 Ligand show cross-species activity. Flt-3 Ligand is expressed as a noncovalently-linked dimer by T cells and bone marrow and thymic fibroblasts. Each 36 kDa chain carries approximately 12 kDa of N- and O-linked carbohydrates. Alternate splicing and proteolytic cleavage of the transmembrane form can generate a soluble 30 kDa fragment that includes the cytokine domain. Alternate splicing of human Flt-3 Ligand also generates membrane-associated isoforms that contain either a truncated cytoplasmic tail or an 85 aa substitution following the cytokine domain. Both transmembrane and soluble Flt-3 Ligand signal through the tyrosine kinase receptor Flt-3/Flk-2. Flt-3 Ligand induces the expansion of monocytes and immature dendritic cells as well as early B cell lineage differentiation. It synergizes with IL-3, GM-CSF, and SCF to promote the mobilization and myeloid differentiation of hematopoietic stem cells. It cooperates with IL-2, -6, -7, and -15 to induce NK cell development and with IL-3, -7, and -11 to induce terminal B cell maturation. Animal studies also show Flt-3 Ligand to reduce the severity of experimentally induced allergic inflammation.
    • Long Name
      fms-like Tyrosine Kinase 3 Ligand
    • Entrez Gene IDs
      2323 (Human); 14256 (Mouse); 493796 (Feline);
    • Alternate Names
      FL; Flt3 ligand; Flt3L; FLT3LG; fms-related tyrosine kinase 3 ligand; SL cytokine;
    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. 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 3 times for a total of 4 washes.

    8. 200 µL Conjugate
    9.   Add 200 µL of Conjugate to each well. Cover with a new plate sealer.
    10.   For Serum & Plasma Samples: Incubate at room temperature for 2 hours.
      For Cell Culture Supernate Samples: Incubate at room temperature for 1.5 hours.
    11.   Aspirate and wash 4 times.

    12. 200 µL Substrate Solution
    13.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.

    14. 100 µL Stop Solution
    15.   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.

    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.

    Showing Results 1 - 10 of 12
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    Sample Type
    1. EphB4 Expressing Stromal Cells Exhibit an Enhanced Capacity for Hematopoietic Stem Cell Maintenance.
      Authors: Nguyen T, Arthur A, Panagopoulos R, Paton S, Hayball J, Zannettino A, Purton L, Matsuo K, Gronthos S
      Stem Cells, 2015;33(9):2838-49.
      Species: Human
      Sample Type: Cell Culture Supernates
    2. Human Flt3L generates dendritic cells from canine peripheral blood precursors: implications for a dog glioma clinical trial.
      Authors: Xiong W, Candolfi M, Liu C
      PLoS ONE, 2010;5(6):e11074.
      Species: Human
      Sample Type: Cell Culture Supernates
    3. Systemic dendritic cell mobilization associated with administration of FLT3 ligand to SIV- and SHIV-infected macaques.
      Authors: Reeves RK, Wei Q, Stallworth J, Fultz PN
      AIDS Res. Hum. Retroviruses, 2009;25(12):1313-28.
      Species: primate-macaca nemestrina (pigtailed macaques)
      Sample Type: Serum
    4. Cerebrospinal fluid Flt3 ligand level in patients with amyotrophic lateral sclerosis.
      Acta Neurol. Scand., 2006;114(3):205-9.
      Species: Human
      Sample Type: CSF
    5. 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
    6. Fms-like tyrosine kinase 3 ligand recruits plasmacytoid dendritic cells to the brain.
      Authors: Curtin JF, King GD, Barcia C, Liu C, Hubert FX, Guillonneau C, Josien R, Anegon I, Lowenstein PR, Castro MG
      J. Immunol., 2006;176(6):3566-77.
      Species: Human
      Sample Type: Serum
    7. Single administration of stem cell factor, FLT-3 ligand, megakaryocyte growth and development factor, and interleukin-3 in combination soon after irradiation prevents nonhuman primates from myelosuppression: long-term follow-up of hematopoiesis.
      Authors: Drouet M, Mourcin F, Grenier N, Leroux V, Denis J, Mayol JF, Thullier P, Lataillade JJ, Herodin F
      Blood, 2004;103(3):878-85.
    8. Inflammatory and anti-glioma effects of an adenovirus expressing human soluble Fms-like tyrosine kinase 3 ligand (hsFlt3L): treatment with hsFlt3L inhibits intracranial glioma progression.
      Authors: Ali S, Curtin JF, Zirger JM, Xiong W, King GD, Barcia C, Liu C, Puntel M, Goverdhana S, Lowenstein PR, Castro MG
      Mol. Ther., 2004;10(6):1071-84.
      Species: Human
      Sample Type: Tissue Homogenates
    9. Fibroblasts from human spleen regulate NK cell differentiation from blood CD34(+) progenitors via cell surface IL-15.
      Authors: Briard D, Brouty-Boye D, Azzarone B, Jasmin C
      J. Immunol., 2002;168(9):4326-32.
      Species: Human
      Sample Type: Cell Culture Supernates
    10. Serum Flt3 ligand variation as a predictive indicator of hematopoietic stem cell mobilization.
      Authors: Haidar JH, Bazarbachi A, Mahfouz R, Haidar HA, Jaafar H, Daher R
      J. Hematother. Stem Cell Res., 2002;11(3):533-8.
      Species: Human
      Sample Type: Serum
    11. 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
    12. CD40-ligand stimulates myelopoiesis by regulating flt3-ligand and thrombopoietin production in bone marrow stromal cells.
      Authors: Solanilla A, Dechanet J, El Andaloussi A, Dupouy M, Godard F, Chabrol J, Charbord P, Reiffers J, Nurden AT, Weksler B, Moreau JF, Ripoche J
      Blood, 2000;95(12):3758-64.
      Species: Human
      Sample Type: Cell Culture Supernates
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