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

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

R&D Systems, part of Bio-Techne | Catalog # 308E-GMP

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

Key Product Details

Source

E. coli

Accession #

P49771.1

Conjugate

Unconjugated

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.

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

Product Datasheets

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COA

SDS

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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:

Manufactured and tested under an ISO 9001:2015 and ISO 13485:2016 certified quality system
Documented processes and QA control of documentation and process changes
Personnel training programs
Raw material testing and vendor qualification/monitoring
Fully validated equipment, processes and test methods
Equipment calibration schedules using a computerized calibration program
Facility maintenance, safety programs and pest control
Material review process for variances
Monitoring of stability over product shelf-life

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 research. The Certificate of Analysis provided contains the following lot specific information:

N-terminal amino acid analysis, SDS-PAGE analysis, mass spectrometry, and endotoxin level (as determined by LAL assay) performed on each bulk QC lot, not on individual bottlings of each QC lot
Post-bottling lot-specific bioassay results (compliance with an established range) and results of microbial testing according to USP
Host Cell Protein testing performed by ELISA
Mycoplasma testing by ribosomal RNA hybridization assay

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, 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.

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.

Please read our complete Animal-Free Statement

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

END USER TERMS OF USE OF PRODUCT

The following terms are offered to you upon your acceptance of these End User Terms of Use of Product. By using this product, you indicate your acknowledgment and agreement to these End User Terms of Use of Product. If you do not agree to be bound by and comply with all of the provisions of these End User Terms of Use of Product, you should contact your supplier of the product and make arrangements to return the product.

We suggest you print and retain a copy of these End User Terms of Use of Product for your records.

The End User is aware that R&D Systems, Inc. sells GMP products for preclinical or clinical ex vivo use and not for in vivo use. The End User further agrees, as a condition of the sale of R&D Systems' GMP products that: a) the End User will not use this GMP Product in any procedure wherein the product may be administered to humans, unless the End User has obtained, or prior to their use will have obtained, an Investigational New Drug (IND) exemption from the FDA and will use the product only in accordance with the protocols of such IND and of the Institutional Review Board overseeing the proposed research, or b) the End User will use the products outside of the United States in accordance with the protocols of research approved by the Institutional Review Board or authorized ethics committee and regulatory agencies to which the End User is subject to in their territory.

R&D Systems, Inc. has the right, at its sole discretion, to modify, add or remove any terms or conditions of these End User Terms of Use without notice or liability to you. Any changes to these End User Terms of Use are effective immediately following the printing of such changes on this product insert. The most recent version of these End User Terms of Use of Product may be found at: RnDSystems.com/Legal.

You agree to review these End User Terms of Use of Product to ensure any subsequent use by you of R&D Systems' GMP Products following changes to these End User Terms of Use of Product constitutes your acceptance of all such changes.

TERMS AND CONDITIONS

The following limitation applies to R&D Systems' warranty and liability for damages: All products are warranted to meet R&D Systems' published specifications when used under normal laboratory conditions.

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For preclinical, or clinical ex vivo use

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