Human Flt-3 Ligand/FLT3L DuoSet ELISA

Catalog #: DY308
5 Citations Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
DY308

Ancillary Products Available

DY008B: DuoSet ELISA Ancillary Reagent Kit 2

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Product Details

Procedure

Citations (5)

Supplemental Products

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Summary Product Features Kit Content Other Reagents Required Data Examples Product Datasheets Preparation and Storage Background

Human Flt-3 Ligand/FLT3L DuoSet ELISA Summary

Assay Type

Solid Phase Sandwich ELISA

Format

96-well strip plate

Sample Volume Required

100 µL

Assay Range

15.6 - 1,000 pg/mL

Sufficient Materials

For fifteen 96-well plates*

Specificity

Please see the product datasheet

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human Flt-3 Ligand. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.

Product Features

Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
Development protocols are provided to guide further assay optimization
Assay can be customized to your specific needs
Economical alternative to complete kits

Kit Content

Capture Antibody
Detection Antibody
Recombinant Standard
Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)

Other Reagents Required

DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.

The components listed above may be purchased separately:

PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na₂HPO₄, 1.5 mM KH₂PO₄, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # WA126), or 0.05% Tween^® 20 in PBS, pH 7.2-7.4

Reagent Diluent: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered

Substrate Solution: 1:1 mixture of Color Reagent A (H₂O₂) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)

Stop Solution: 2 N H₂SO₄ (Catalog # DY994)

Microplates: R&D Systems (Catalog # DY990)

Plate Sealers: ELISA Plate Sealers (Catalog # DY992)

Scientific Data

N/A

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Human Flt-3 Ligand ELISA Standard Curve

Product Datasheets

Product Datasheet

COA

SDS

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 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

Assay Procedure

GENERAL ELISA PROTOCOL

Plate Preparation

Dilute the Capture Antibody to the working concentration in PBS without carrier protein. Immediately coat a 96-well microplate with 100 μL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 μL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

Assay Procedure

Add 100 μL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
Repeat the aspiration/wash as in step 2 of Plate Preparation.
Add 100 μL of the Detection Antibody, diluted in Reagent Diluent, to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
Repeat the aspiration/wash as in step 2 of Plate Preparation.
Add 100 μL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
Repeat the aspiration/wash as in step 2.
Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.

Citations for Human Flt-3 Ligand/FLT3L DuoSet ELISA

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.

5 Citations: Showing 1 - 5
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A new cytokine-based dynamic stratification during induction is highly predictive of survivals in acute myeloid leukemia
Authors: P Peterlin, J Gaschet, T Guillaume, A Garnier, M Eveillard, A Le Bourgeo, M Cherel, C Debord, Y Le Bris, O Theisen, C Godon, B Mahé, V Dubruille, S Wuilleme, C Touzeau, T Gastinne, N Blin, A Lok, B Tessoulin, S Le Gouill, P Moreau, MC Béné, P Chevallier
Cancer Medicine, 2020-12-25;0(0):.
Species: Human
Sample Types: Serum
FLT3 ligand plasma levels have no impact on outcomes after allotransplant in acute leukemia
Authors: P Peterlin, J Gaschet, T Guillaume, A Garnier, M Eveillard, A Le Bourgeo, M Cherel, C Debord, Y Le Bris, O Theisen, B Mahé, V Dubruille, C Godon, N Robillard, S Wuilleme, C Touzeau, T Gastinne, N Blin, A Lok, A Bonnet, S Le Gouill, P Moreau, MC Béné, P Chevallier
Cytokine, 2019-04-28;120(0):85-87.
Species: Human
Sample Types: Plasma
FLT3 ligand plasma levels in acute myeloid leukemia
Authors: P Peterlin, J Gaschet, T Guillaume, A Garnier, M Eveillard, A Le Bourgeo, M Cherel, C Debord, Y Le Bris, O Theisen, B Mahé, V Dubruille, C Godon, N Robillard, S Wuillem, C Touzeau, T Gastinne, N Blin, A Lok, A Bonnet, S Le Gouill, P Moreau, MC Béné, P Chevallier
Haematologica, 2019-01-10;0(0):.
Species: Human
Sample Types: Plasma
Activation of Fms-like tyrosine kinase 3 signaling enhances survivin expression in a mouse model of rheumatoid arthritis.
Authors: Andersson S, Svensson M, Erlandsson M, Dehlin M, Andersson K, Bokarewa M
PLoS ONE, 2012-10-17;7(10):e47668.
Species: Human
Sample Types: Plasma
Barley as a green factory for the production of functional Flt3 ligand.
Authors: Erlendsson LS, Muench MO, Hellman U, Hrafnkelsdottir SM, Jonsson A, Balmer Y, Mantyla E, Orvar BL
Biotechnol J, 2010-02-01;5(2):163-71.
Species: Human
Sample Types: Recombinant Protein