Recombinant Human Flt-3 Ligand/FLT3L GMP Protein, CF Summary
Product Specifications
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).
Thr27-Ala181
Produced in an animal-free laboratory. Manufactured and tested under cGMP guidelines
Analysis
Met-Thr27-Gln-Asp-(Cys)-Ser-Phe-Gln-His-Ser and
Thr27-Gln-Asp-(Cys)-Ser-Phe-Gln-His-Ser-Pro
Product Datasheets
Carrier Free
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.
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.
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.
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Scientific Data
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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.
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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.
Reconstitution Calculator
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).
- 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.
Manufacturing Specifications
GMP ProteinsR&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
- 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
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.
Production
- All molecular biology procedures use animal-free media and dedicated labware.
- Dedicated fermentors are utilized in committed animal-free areas.
- Protein purification columns are animal-free.
- Bulk proteins are filtered using animal-free filters.
- Purified proteins are stored in animal-free containers.
Product Specific Notices
Full terms and conditions of sale can be found online in the Protein Sciences Segment T&Cs at: Terms & Conditions.FAQs
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