GMP

Recombinant Human Flt-3 Ligand/FLT3L GMP Protein, CF

Name: Recombinant Human Flt-3 Ligand/FLT3L GMP Protein, CF
Brand: R&D Systems
SKU: 308E-GMP

R&D Systems | Catalog # 308E-GMP

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Product Specifications
Scientific Data
Preparation & Storage
Calculators
Background
Product Documents
Manufacturing Specifications
Specific Notices
Research Areas

Key Product Details

Source

E. coli

Accession Number

P49771.1

Applications

Bioactivity

View all Flt-3 Ligand/FLT3L Proteins and Enzymes »

Product Specifications

Source

E. coli-derived human Flt-3 Ligand/FLT3L protein
Thr27-Ala181
Produced in an animal-free laboratory. Manufactured and tested under cGMP guidelines

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-Thr27-Gln-Asp-(Cys)-Ser-Phe-Gln-His-Ser and

Thr27-Gln-Asp-(Cys)-Ser-Phe-Gln-His-Ser-Pro

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.
The specific activity of Recombinant Human Flt-3 Ligand/FLT3L is >5.0 x 10⁵ units/mg, which is calibrated against the human Flt-3 Ligand WHO Standard (NIBSC code: 96/532).

Host Cell Protein

< 0.5 ng per μg of protein when tested by ELISA.

Mycoplasma

Negative for Mycoplasma.

Host Cell DNA

< 0.0010 ng per µg of protein when tested by PCR.

Scientific Data Images for Recombinant Human Flt-3 Ligand/FLT3L GMP Protein, CF

Recombinant Human Flt-3 Ligand/FLT3L GMP Protein Bioactivity

GMP-grade Recombinant Human Flt-3 Ligand/FLT3L (Catalog # 308E-GMP) stimulates cell proliferation in the BaF3 mouse pro-B cell line transfected with mouse Flt-3. The ED₅₀ for this effect is 0.2-1 ng/mL. Three independent lots were tested for activity and plotted on the same graph to show lot-to-lot consistency of GMP Flt‑3 Ligand/FLT3L.

Recombinant Human Flt-3 Ligand/FLT3L GMP Protein SDS-PAGE

2 μg/lane of GMP-grade Recombinant Human Flt-3 Ligand/FLT3L (Catalog # 308E-GMP) 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

308E-GMP

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 with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage	Use a manual defrost freezer and avoid repeated freeze-thaw cycles. A minimum of 12 months when stored at ≤ -20 °C as supplied. Refer to lot specific COA for the Use by Date. 1 month, 2 to 8 °C under sterile conditions after reconstitution. 3 months, ≤ -20 °C under sterile conditions after reconstitution.

Calculators

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.

Volume (to add to vial)

Mass (in vial)

Desired Reconstitution Concentration

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).

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

Alternate Names

FLG3L, Flt3L, FLT3LG

Entrez Gene IDs

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

Gene Symbol

FLT3LG

UniProt

P49771.1

Additional Flt-3 Ligand/FLT3L Products

Product Documents for Recombinant Human Flt-3 Ligand/FLT3L GMP Protein, CF

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Certificate of Analysis

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Manufacturing Specifications

GMP Proteins
R&D Systems, a Bio-Techne Brand's GMP proteins are produced according to relevant sections of the following documents: USP Chapter 1043, Ancillary Materials for Cell, Gene and Tissue-Engineered Products and Eu. Ph. 5.2.12, Raw Materials of Biological Origin for the Production of Cell-based and Gene Therapy Medicinal Products.

R&D Systems' quality focus includes:

Designed, manufactured and tested under an ISO 9001:2015 and ISO 13485:2016 certified quality system
Documented and controlled manufacturing process
Control of documentation and process changes by QA
Personnel training programs
Raw material inspection and vendor qualification/monitoring program
Validated equipment, processes and test methods
Equipment calibration and maintenance schedules using a Regulatory Asset Manager
Facility/Utilities maintenance, contamination controls, safety and pest control programs
Material review process for variances
Robust product stability program following relevant ICH guidelines

R&D Systems strives to provide our customers with the analytical characteristics of each product so that customers may determine whether our products are appropriate for their application. Each product is provided with a lot-specific Certificate of Analysis that contains the product's specifications and test results. Quality control testing may include, but is not limited to:

N-terminal amino acid analysis
SDS-PAGE purity analysis
Molecular weight analysis via mass spectrometry
Endotoxin assessment per USP <85> and Ph. Eur. 2.6.14 guidelines
Bioassay analysis
Microbial testing per USP <71> and Ph. Eur. 2.6.1 guidelines
Host cell protein assessment
Host cell DNA assessment
Mycoplasma assessment

Additional testing and documentation requested by the customer can be arranged at an additional cost.

Production records and facilities are available for examination by appropriate personnel on-site at R&D Systems in Minneapolis and St. Paul, Minnesota USA.

R&D Systems sells GMP grade products for preclinical or clinical ex vivo use. They are not for in vivo use. Please read the following End User Terms prior to using this product.

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.

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