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Human Flt-3 Ligand/FLT3L Quantikine ELISA Kit

R&D Systems | Catalog # DFK00

R&D Systems
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Key Product Details

Assay Length

4 hours or 4.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (50 µL), Serum (50 µL), EDTA Plasma (50 µL), Heparin Plasma (50 µL), Citrate Plasma (50 µL)

Sensitivity

7 pg/mL

Assay Range

15.6-1000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma)
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Product Summary for Human Flt-3 Ligand/FLT3L Quantikine ELISA Kit

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.

Product Specifications

Assay Type

Solid Phase Sandwich ELISA

Format

96-well strip plate

Measurement

Quantitative ELISA

Detection Method

Colorimetric - 450nm (TMB)

Conjugate

HRP

Species

Human

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.

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

Citrate Plasma, EDTA Plasma, Heparin Plasma, Serum

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 for Human Flt-3 Ligand/FLT3L Quantikine ELISA Kit

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 Images for Human Flt-3 Ligand/FLT3L Quantikine ELISA Kit

Human Flt-3 Ligand ELISA Cell Culture Supernate Standard Curve

Human Flt-3 Ligand ELISA Cell Culture Supernate Standard Curve

Human Flt-3 Ligand ELISA Serum/Plasma Standard Curve

Human Flt-3 Ligand ELISA Serum/Plasma Standard Curve

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: Flt-3 Ligand/FLT3L

Flt-3 Ligand, also known as FLT3L, is a cytokine that promotes the differentiation or expansion of multiple hematopoietic cell lineages including monocytes, immature dendritic cells, and early B cell lineage cells. It synergizes with other cytokines to promote the myeloid differentiation and mobilization of hematopoietic stem cells, to induce NK cell development, and to induce terminal B cell maturation. Flt-3 Ligand/FLT3L is expressed as a dimer by T cells and bone marrow and thymic fibroblasts. Both transmembrane and soluble forms of Flt-3 Ligand/FLT3L signal through the tyrosine kinase receptor Flt-3/Flk-2.

Long Name

fms-like Tyrosine Kinase 3 Ligand

Alternate Names

FLG3L, Flt3L, FLT3LG

Entrez Gene IDs

2323 (Human); 14256 (Mouse); 102125470 (Cynomolgus Monkey); 493796 (Feline)

Gene Symbol

FLT3LG

Additional Flt-3 Ligand/FLT3L Products

Product Documents for Human Flt-3 Ligand/FLT3L Quantikine ELISA Kit

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human Flt-3 Ligand/FLT3L Quantikine ELISA Kit

For research use only

⚠ 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.

Citations for Human Flt-3 Ligand/FLT3L Quantikine ELISA Kit

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  • Human Flt-3 Ligand Quantikine ELISA Kit
    Name: Alisha Freeman
    Sample Tested: Murine mesenteric lymph node
    Verified Customer | Posted 07/12/2018
  • Human Flt-3 Ligand Quantikine ELISA Kit
    Name: Alisha Freeman
    Sample Tested: CHO Chinese hamster ovary cell line
    Verified Customer | Posted 12/21/2017
  • Human Flt-3 Ligand Quantikine ELISA Kit
    Name: Robert Keith
    Sample Tested: Normal human serum samples
    Verified Customer | Posted 12/07/2017
    ELISA detected all tested human normal serum samples with range of 54 to 111 pg/mL.
    Human Flt-3 Ligand/FLT3L Quantikine ELISA Kit DFK00

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Protocols

View specific protocols for Human Flt-3 Ligand/FLT3L Quantikine ELISA Kit (DFK00):

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.

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FAQs

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