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Human Flt-3 Ligand/FLT3L ELISA Kit - Quantikine

Catalog #: DFK00
21 Citations 3 Reviews Datasheet / COA / SDS
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Human Flt-3 Ligand ELISA Cell Culture Supernate Standard Curve
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Citations (21)
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Summary Precision Recovery Linearity Data Examples Product Datasheets Preparation and Storage Background Related Research Areas

Human Flt-3 Ligand/FLT3L Quantikine ELISA Kit Summary

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.

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.

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

Cell Culture Supernates

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean (pg/mL) 65.7 214 421 72.8 224 429
Standard Deviation 1.2 4.1 9.3 6.7 13.8 30.5
CV% 1.8 1.9 2.2 9.2 6.2 7.1

Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean (pg/mL) 69.5 230 453 78.2 241 462
Standard Deviation 1.9 5.2 6.2 8.7 14.9 33.1
CV% 2.7 2.3 1.4 11.1 6.2 7.2

Recovery

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

Linearity

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 ELISA Linearity

Scientific Data

N/A Human Flt-3 Ligand ELISA Cell Culture Supernate Standard Curve View Larger

Human Flt-3 Ligand ELISA Cell Culture Supernate Standard Curve

N/A Human Flt-3 Ligand ELISA Serum/Plasma Standard Curve View Larger

Human Flt-3 Ligand ELISA Serum/Plasma 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.
Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: Flt-3 Ligand/FLT3L

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); 102125470 (Cynomolgus Monkey); 493796 (Feline)
Alternate Names:
FL; FLG3L; Flt3 ligand; Flt-3 Ligand; Flt3L; FLT3LG; fms-related tyrosine kinase 3 ligand; SL cytokine
⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.

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
  3.   Add 100 µL of Assay Diluent to each well.
    4. 50 µL Standard, Control, or Sample
  4.   Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
  5.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
    6. 200 µL Conjugate
  6.   Add 200 µL of Conjugate to each well. Cover with a new plate sealer.
  7.   For Serum & Plasma Samples: Incubate at room temperature for 2 hours.
    For Cell Culture Supernate Samples: Incubate at room temperature for 1.5 hours.
  8.   Aspirate and wash 4 times.
    9. 200 µL Substrate Solution
  9.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.
    10. 100 µL Stop Solution
  10.   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.

Citations for Human Flt-3 Ligand/FLT3L Quantikine 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.

21 Citations: Showing 1 - 10
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  1. Elevated Flt3L Predicts Long-Term Survival in Patients with High-Grade Gastroenteropancreatic Neuroendocrine Neoplasms
    Authors: KM Detjen, R Otto, Y Giesecke, L Geisler, P Riemer, H Jann, C Grötzinger, C Sers, A Pascher, T Lüdde, U Leser, B Wiedenmann, M Sigal, F Tacke, C Roderburg, L Hammerich
    Cancers, 2021-09-04;13(17):.
    Species: Human
    Sample Types: Serum
  2. Reduced circulating dendritic cells in acute Plasmodium knowlesi and Plasmodium falciparum malaria despite elevated plasma Flt3 ligand levels
    Authors: JR Loughland, T Woodberry, D Oyong, KA Piera, FH Amante, BE Barber, MJ Grigg, T William, CR Engwerda, NM Anstey, JS McCarthy, MJ Boyle, G Minigo
    Malaria Journal, 2021-02-16;20(1):97.
    Species: Human
    Sample Types: Plasma
  3. FLT3 stop mutation increases FLT3 ligand level and risk of autoimmune thyroid disease
    Authors: S Saevarsdot, TA Olafsdotti, EV Ivarsdotti, GH Halldorsso, K Gunnarsdot, A Sigurdsson, A Johannesso, JK Sigurdsson, T Juliusdott, SH Lund, AO Arnthorsso, EL Styrmisdot, J Gudmundsso, GM Grondal, K Steinsson, L Alfredsson, J Askling, R Benediktss, R Bjarnason, AJ Geirsson, B Gudbjornss, H Gudjonsson, H Hjaltason, AB Hreidarsso, L Klareskog, I Kockum, H Kristjansd, TJ Love, BR Ludviksson, T Olsson, PT Onundarson, KB Orvar, L Padyukov, B Sigurgeirs, V Tragante, K Bjarnadott, T Rafnar, G Masson, P Sulem, DF Gudbjartss, P Melsted, G Thorleifss, GL Norddahl, U Thorsteins, I Jonsdottir, K Stefansson
    Nature, 2020-06-24;0(0):.
    Species: Human
    Sample Types: Plasma
  4. Effect of Fms-like tyrosine kinase 3 (FLT3) ligand (FL) on antitumor activity of gilteritinib, a FLT3 inhibitor, in mice xenografted with FL-overexpressing cells
    Authors: T Kawase, T Nakazawa, T Eguchi, H Tsuzuki, Y Ueno, Y Amano, T Suzuki, M Mori, T Yoshida
    Oncotarget, 2019-10-22;10(58):6111-6123.
    Species: Human
    Sample Types: Cell Culture Supernates
  5. HIV-1 Tat protein inhibits the hematopoietic support function of human bone marrow mesenchymal stem cells
    Authors: Y Yuan, C Zhou, Q Yang, S Ma, X Wang, X Guo, Y Ding, J Tang, Y Zeng, D Li
    Virus Res., 2019-09-12;273(0):197756.
    Species: Human
    Sample Types: Cell Culture Supernates
  6. Mesenchymal stem cell therapy induces FLT3L and CD1c+ dendritic cells in systemic lupus erythematosus patients
    Authors: X Yuan, X Qin, D Wang, Z Zhang, X Tang, X Gao, W Chen, L Sun
    Nat Commun, 2019-06-07;10(1):2498.
    Species: Human
    Sample Types: Cell Culture Supernates
  7. Systemic immune response induced by oxaliplatin-based neoadjuvant therapy favours survival without metastatic progression in high-risk rectal cancer
    Authors: E Kalanxhi, S Meltzer, JV Schou, FO Larsen, S Dueland, K Flatmark, BV Jensen, KH Hole, T Seierstad, KR Redalen, DL Nielsen, AH Ree
    Br. J. Cancer, 2018-04-26;118(10):1322-1328.
    Species: Human
    Sample Types: Serum
  8. Differential expression of cell cycle and WNT pathway-related genes accounts for differences in the growth and differentiation potential of Wharton's jelly and bone marrow-derived mesenchymal stem cells
    Authors: AK Batsali, C Pontikoglo, D Koutroulak, KI Pavlaki, A Damianaki, I Mavroudi, K Alpantaki, E Kouvidi, G Kontakis, HA Papadaki
    Stem Cell Res Ther, 2017-04-26;8(1):102.
    Species: Human
    Sample Types: Cell Culture Supernates
  9. Timely follow-up of a GATA2 deficiency patient allows successful treatment
    Authors: S Ciullini M, M Vignoli, G Colarusso, F Tucci, M Veltroni, S Frenos, V Tintori, M Aricò, V Bigley, M Collin, C Favre, E Gambineri
    J. Allergy Clin. Immunol, 2016-07-29;138(5):1480-1483.e4.
    Species: Human
    Sample Types: Serum
  10. 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, 2015-06-23;33(9):2838-49.
    Species: Human
    Sample Types: Cell Culture Supernates
  11. 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, 2010-06-11;5(6):e11074.
    Species: Human
    Sample Types: Cell Culture Supernates
  12. 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, 2009-12-01;25(12):1313-28.
    Species: Primate - Macaca nemestrina (Southern Pig-tailed Macaque)
    Sample Types: Serum
  13. 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, 2006-11-15;119(10):2313-21.
    Species: Human
    Sample Types: Cell Culture Supernates
  14. Cerebrospinal fluid Flt3 ligand level in patients with amyotrophic lateral sclerosis.
    Acta Neurol. Scand., 2006-09-01;114(3):205-9.
    Species: Human
    Sample Types: CSF, Serum
  15. 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., 2006-03-15;176(6):3566-77.
    Species: Human
    Sample Types: Serum
  16. 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., 2004-12-01;10(6):1071-84.
    Species: Human
    Sample Types: Tissue Homogenates
  17. 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, 2003-10-02;103(3):878-85.
    Species: Primate - Macaca fascicularis (Crab-eating Monkey or Cynomolgus Macaque)
    Sample Types: Plasma
  18. 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., 2002-05-01;168(9):4326-32.
    Species: Human
    Sample Types: Cell Culture Supernates
  19. 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., 2001-06-01;29(6):736-45.
    Species: Human
    Sample Types: Cell Culture Supernates
  20. 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, 2000-06-15;95(12):3758-64.
    Species: Human
    Sample Types: Cell Culture Supernates

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Human Flt-3 Ligand Quantikine ELISA Kit
By Alisha Freeman on 07/12/2018
Sample Tested: Murine mesenteric lymph node
Human Flt-3 Ligand Quantikine ELISA Kit
By Alisha Freeman on 12/21/2017
Sample Tested: CHO Chinese hamster ovary cell line
Human Flt-3 Ligand Quantikine ELISA Kit
By Robert Keith on 12/07/2017
Sample Tested: Normal human serum samples

ELISA detected all tested human normal serum samples with range of 54 to 111 pg/mL.

Human Flt-3 Ligand Quantikine ELISA Kit DFK00