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AB254507

Human Flt-3 ligand ELISA Kit

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Human Flt-3 ligand ELISA Kit is a single-wash 90-min Simplestep used to quantify Human Flt-3 ligand with a sensitivity of 2 pg/ml. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.

- Colorimetric Sandwich ELISA - 450 nm readout : works on any standard plate reader
- Different formats for different needs: 10x96 plates for bulk orders and 384-well plate for higher throughput
- Design your own immunoassay: we also offer the conjugation-ready antibody pair

View Alternative Names

Fms-related tyrosine kinase 3 ligand, Flt3 ligand, Flt3L, SL cytokine, FLT3LG

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ELISA - Human Flt-3 ligand ELISA Kit (AB254507)
  • ELISA

Supplier Data

ELISA - Human Flt-3 ligand ELISA Kit (AB254507)
Sandwich ELISA - Human Flt-3 ligand ELISA Kit (AB254507)
  • sELISA

Lab

Sandwich ELISA - Human Flt-3 ligand ELISA Kit (AB254507)

Interpolated concentration of native Flt-3 Ligand was measured in duplicate at different sample concentrations in 96-well vs. 384-well plates. Undiluted samples are 50% human serum. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). Sample dilutions are made in Sample Diluent NS.

Sandwich ELISA - Human Flt-3 ligand ELISA Kit (AB254507)
  • sELISA

Supplier Data

Sandwich ELISA - Human Flt-3 ligand ELISA Kit (AB254507)

Human Fit-3 Ligand standard curve comparison.

Standard Curve comparison between human Fit-3 Ligand SimpleStep ELISA kit and traditional ELISA kit from leading competitor. SimpleStep ELISA kit shows increased sensitivity.

Sandwich ELISA - Human Flt-3 ligand ELISA Kit (AB254507)
  • sELISA

Supplier Data

Sandwich ELISA - Human Flt-3 ligand ELISA Kit (AB254507)

Interpolated concentrations of native Flt-3 Ligand in human serum and plasma samples.

The concentrations of Flt-3 Ligand were measured in duplicates, interpolated from the Flt-3 Ligand standard curves and corrected for sample dilution. Undiluted samples are as follows : serum 50%, plasma (citrate) 50%, plasma (heparin) 50%, and plasma (EDTA) 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Flt-3 Ligand concentration was determined to be 65.35 pg/mL in neat serum, 57.01 pg/mL in neat plasma (citrate), 54.11 pg/mL in neat plasma (heparin), and 64.57 pg/mL in neat plasma (EDTA).

Sandwich ELISA - Human Flt-3 ligand ELISA Kit (AB254507)
  • sELISA

Lab

Sandwich ELISA - Human Flt-3 ligand ELISA Kit (AB254507)

Example of human Flt-3 Ligand standard curve in 96-well vs. 384-well plate. Background-subtracted data values (mean +/- SD) are graphed.

Sandwich ELISA - Human Flt-3 ligand ELISA Kit (AB254507)
  • sELISA

Supplier Data

Sandwich ELISA - Human Flt-3 ligand ELISA Kit (AB254507)

Interpolated concentrations of native Flt-3 Ligand in human PBMC cell culture supernatant samples. Human PBMC were cultured in the presence (stimulated) or absence (unstimulated) of 1.5% PHA-M.

Human peripheral blood mononuclear cells (PBMC) were cultured in the presence (stimulated) or absence (unstimulated) of 1.5% PHA-M for 36 hours. The concentrations of Flt-3 Ligand were measured in duplicates, interpolated from the Flt-3 Ligand standard curves and corrected for sample dilution. Undiluted samples are as follows : stimulated PBMC supernatant 100% and unstimulated PBMC supernatant 100%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Flt-3 Ligand concentration was determined to be 22.72 pg/mL in neat PHA-M stimulated PBMC cell culture supernatant, 3.742 pg/mL in neat unstimulated PBMC cell culture supernatant, and undetectable in naïve cell culture media (data not shown).

Sandwich ELISA - Human Flt-3 ligand ELISA Kit (AB254507)
  • sELISA

Supplier Data

Sandwich ELISA - Human Flt-3 ligand ELISA Kit (AB254507)

Serum from ten individual healthy human female donors was measured in duplicate.

Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Flt-3 Ligand concentration was determined to be 63.87 pg/mL with a range of 26.63 – 105.1 pg/mL.

Sandwich ELISA - Human Flt-3 ligand ELISA Kit (AB254507)
  • sELISA

Supplier Data

Sandwich ELISA - Human Flt-3 ligand ELISA Kit (AB254507)

Example of human Flt-3 Ligand standard curve in Sample Diluent NS.

The Flt-3 Ligand standard curve was prepared as described in Section 10. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.

Key facts

Detection method

Colorimetric

Sample types

Heparin Plasma, Citrate plasma, Cell culture supernatant, Serum, EDTA Plasma

Reacts with

Human

Assay type

Sandwich (quantitative)

Sensitivity

= 2 pg/mL

Range

4.7 - 300 pg/mL

Assay time

1h 30m

Assay Platform

Pre-coated microplate (12 x 8 well strips)

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "sELISA": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }
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Product details

Human Flt-3 ligand ELISA Kit ab254507 is a rapid single-wash 90-min Sandwich ELISA to measure Human Flt-3 ligand in cell culture supernatant, citrate plasma, EDTA plasma, heparin plasma, serum. This SimpleStep sensitivity is 2 pg/mL.

How the assay works

Human Flt-3 ligand SimpleStep ELISA®employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details.

Assay Specificity

Our SimpleStep ELISA® kits use recombinant monoclonal antibodies rigorously validated to ensure the highest level of consistency and reproducibility, improved sensitivity and specificity and ease of scalability and security of supply.
Please refer to our protocol booklet for more details.

Human Flt-3 ligand ELISA Kit ab254507 protocol summary

1. Mix: add samples/standards to the wells together with the capture and detector antibody cocktail. Incubate 1 hr at room temperature
2. Wash
3. Add TMB development solution - incubate for 10 min
4. Add Stop solution
5. Read the results on a plate reader at 450 nm

Save your precious samples

Our 384-well Human Flt-3 ligand ELISA Kit only requires a maximum of 12.5µL sample

Design your own immunoassay

We offer the antibody pair used in this kit in a BSA and Azide-free format, ready for conjugation:

- Anti-Flt3 ligand/Flt3L antibody [EPR22288-52] - BSA and Azide free (Capture) ab259726
- Anti-Flt3 ligand/Flt3L antibody [EPR22288-201] - BSA and Azide free (Detector) ab259726

Flt-3 Ligand binds to and activates Flt-3. Flt-3 Ligand works synergistically with other CSFs and interleukins to stimulate growth and differentiation of early hematopoietic cells. The predominant form of Flt-3 Ligand is the biologically active transmembrane form which is proteolytically cleaved to generate a soluble form that is also biologically active. Alternative spliced secreted soluble variant of Flt-3 Ligand has also been identified.

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Precision

[ { "reproducibilityType": "Inter", "sample": "Serum", "replicates": 3, "mean": null, "standardDeviation": null, "coefficientOfVariability": "4.2" }, { "reproducibilityType": "Intra", "sample": "Serum", "replicates": 8, "mean": null, "standardDeviation": null, "coefficientOfVariability": "4.1" } ]
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Recovery

[ { "sample": "Cell culture supernatant", "range": "94 - 97 %", "average": "= 95" }, { "sample": "Serum", "range": "93 - 103 %", "average": "= 99" }, { "sample": "EDTA Plasma", "range": "87 - 97 %", "average": "= 94" }, { "sample": "Heparin Plasma", "range": "93 - 101 %", "average": "= 98" }, { "sample": "Citrate plasma", "range": "93 - 101 %", "average": "= 97" } ]
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What's included?

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Properties and storage information

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
Storage information
+4°C
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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

The Flt3 ligand (Flt3L) also known as Flt3-L Flt3 ligand protein or flt-3 ligand protein is a small cytokine with a molecular mass of approximately 18 to 19 kDa. It acts as a growth factor by binding to the Flt3 receptor tyrosine kinase on the surface of hematopoietic progenitor cells. Flt3L is expressed in various tissues including the bone marrow where it plays a significant role in hematopoiesis. It is critical in dendritic cell and B cell development impacting the immune response mechanism.
Biological function summary

The Flt3 ligand plays an essential role in the immune system. It is a monomeric cytokine that induces proliferation of early lineage cells within the hematopoietic system. Flt3L does not function as part of a complex but acts independently to stimulate the proliferation differentiation and survival of hematopoietic cells. By interacting directly with its receptor it significantly influences the innate and adaptive immune systems through the maturation of dendritic and B cells.

Pathways

The Flt3 ligand is involved in hematopoietic signaling pathways primarily the PI3K-Akt signaling pathway. This pathway is essential for cell survival and growth where Flt3L aids in transmitting growth signals necessary for hematopoietic cells. Flt3L also interacts with proteins such as FLT3 receptor and influences the downstream signaling cascade affecting cell fate decisions in hematopoiesis and immune cell maturation.

Flt3 ligand is connected to leukemia particularly acute myeloid leukemia (AML) due to dysregulation of the FLT3 signaling pathway. Mutations in the FLT3 receptor can lead to uncontrolled cell proliferation contributing to tumorigenesis. Additionally Flt3L plays a role in autoimmune disorders by influencing immune cell development. It is closely connected with the FLT3 receptor and other tyrosine kinases which when mutated can exacerbate disease pathology through abnormal cell signaling and proliferation.
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Product protocols

For this product, it's our understanding that no specific protocols are required. You can visit:
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Target data

Stimulates the proliferation of early hematopoietic cells by activating FLT3. Synergizes well with a number of other colony stimulating factors and interleukins.
See full target information FLT3LG
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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions
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