Recombinant Human Flt-3 Ligand/FLT3L Animal-Free Protein

Name: Recombinant Human Flt-3 Ligand/FLT3L Animal-Free Protein
Brand: rnd
SKU: AFL308E

Catalog # AFL308E

Also available, GMP FLT3L Protein, Catalog # 308E-GMP! Animal-free and manufactured under GMP guidelines for therapeutic manufacturing applications.

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Key Product Details

Source:	E. coli
Accession #:	P49771.1
Applications:	Bioactivity

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Product Specifications

Source	E. coli-derived human Flt-3 Ligand/FLT3L protein Thr27-Ala181 Produced using non-animal reagents in an animal-free laboratory.
Purity	>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin level	<0.10 EU per 1 μg of the protein by the LAL method.
N-terminal sequence Analysis	Met and Thr27
Predicted Molecular Mass	18 kDa
SDS-PAGE	15-17 kDa, under reducing conditions
Activity	Measured in a cell proliferation assay using BaF3 mouse pro‑B cells transfected with mouse Flt-3. The ED₅₀ for this effect is 0.2-1 ng/mL

Recombinant Human Flt-3 Ligand/FLT3L Animal-Free Protein Scientific Data Examples

	Recombinant Human Flt-3 Ligand/FLT3L Animal-Free Protein Bioactivity Recombinant Human Flt-3 Ligand/FLT3L Animal-Free Protein (Catalog # AFL308E) stimulates cell proliferation in the BaF3 mouse pro-B cells transfected with mouse Flt-3. The ED₅₀ for this effect is 0.2-1 ng/mL.
	Recombinant Human Flt-3 Ligand/FLT3L Animal-Free Protein SDS-PAGE 2 μg/lane of Animal-Free™ Recombinant Human Flt-3 Ligand/FLT3L (Catalog # AFL308E) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® blue staining, showing bands at 15-17 kDa and 12-14 kDa, respectively.

Formulation, Preparation and Storage

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

AFL308E

Formulation	Lyophilized from a 0.2 μm filtered solution in PBS.
Reconstitution	Reconstitute at 100-500 μg/mL in PBS.
Shipping	The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage:	Use a manual defrost freezer and avoid repeated freeze-thaw cycles. 12 months from date of receipt, -20 to -70 °C as supplied. 1 month, 2 to 8 °C under sterile conditions after reconstitution. 3 months, -20 to -70 °C under sterile conditions after reconstitution.

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: Flt-3 Ligand/FLT3L

Flt‑3 Ligand, also known as FLT3L, is an alpha-helical cytokine that promotes the differentiation of multiple hematopoietic cell lineages (1-3). 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 (4-7). Within the ECD, human Flt‑3 Ligand shares 71% and 65% aa sequence identity with mouse and rat Flt‑3 Ligand, respectively (4-6). The human and mouse Flt‑3 Ligand proteins show cross-species activity. Flt-3 Ligand is also structurally related to M-CSF and SCF. Flt-3 Ligand is widely expressed in various human and mouse tissues. It is expressed as a noncovalently-linked dimer by T cells and bone marrow and thymic fibroblasts (1, 8). Each 36 kDa chain of the Flt-3 Ligand dimer carries approximately 12 kDa of N- and O-linked carbohydrates (8). Alternate splicing and proteolytic cleavage of the transmembrane form of the Flt-3 Ligand protein can generate a soluble 30 kDa fragment that includes the cytokine-like domain (4, 8). 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-like domain in the ECD of the Flt-3 Ligand protein (4, 5, 8). Both transmembrane and soluble forms of Flt‑3 Ligand signal through the tyrosine kinase receptor Flt-3/Flk-2 (3, 4, 6, 7). Flt‑3 Ligand induces the expansion of monocytes and immature dendritic cells as well as early B cell lineage differentiation (2, 9). Additionally, Flt-3 Ligand synergizes with IL-3, GM-CSF, and SCF to promote the mobilization and myeloid differentiation of hematopoietic stem cells (4-6). Flt-3 Ligand also cooperates with IL-2, IL-6, IL-7, and IL-15 to induce NK cell development and with IL-3, IL-7, and IL-11 to induce terminal B cell maturation (1, 10). Animal studies show that Flt‑3 Ligand reduces the severity of experimentally induced allergic inflammation (11).

Item	Value
References	Wodnar-Filipowicz, A. (2003) News Physiol. Sci. 18:247. Dong, J. et al. (2002) Cancer Biol. Ther. 1:486. Gilliland, D.G. and J.D. Griffin (2002) Blood 100:1532. Hannum, C. et al. (1994) Nature 368:643. Lyman, S.D. et al. (1994) Blood 83:2795. Lyman, S.D. et al. (1993) Cell 75:1157. Savvides, S.N. et al. (2000) Nat. Struct. Biol. 7:486. McClanahan, T. et al. (1996) Blood 88:3371. Diener, K.R. et al. (2008) Exp. Hematol. 36:51. Farag, S.S. and M.A. Caligiuri (2006) Blood Rev. 20:123. Edwan, J.H. et al. (2004) J. Immunol. 172:5016.
Long Name	fms-like Tyrosine Kinase 3 Ligand
Entrez Gene IDs	2323 (Human); 14256 (Mouse); 102125470 (Cynomolgus Monkey); 493796 (Feline)
Alternate Names	FL, FLG3L, FLT3LG, Flt-3 Ligand, Flt3 ligand, Flt3L, SL cytokine, fms-related tyrosine kinase 3 ligand

Manufacturing Specifications

Animal-Free Manufacturing Conditions
Our dedicated controlled-access animal-free laboratories ensure that at no point in production are the products exposed to potential contamination by animal components or byproducts. Every stage of manufacturing is conducted in compliance with R&D Systems' stringent Standard Operating Procedures (SOPs). Production and purification procedures use equipment and media that are confirmed animal-free.

Production

All molecular biology procedures use animal-free media and dedicated labware.
Dedicated fermentors are utilized in committed animal-free areas.

Purification

Protein purification columns are animal-free.
Bulk proteins are filtered using animal-free filters.
Purified proteins are stored in animal-free containers in a dedicated cold storage room.

Quality Assurance

Low Endotoxin Level.
No impairment of biological activity.
High quality product obtained under stringent conditions.
For ex vivo research or bioproduction, additional documentation can be provided.

Please read our complete Animal-Free Statement

Citations

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