Recombinant Human Flt-3 Ligand/FLT3L (E. coli) Protein, CF Summary
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.
|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.
Recombinant Human Flt-3 Ligand/FLT3L(E. coli) (Catalog # 308-FKE) stimulates cell proliferation in the BaF3 mouse pro-B cell line transfected with mouse Flt-3. The ED50for this effect is 0.2-1 ng/mL.
2 μg/lane of Recombinant Human Flt-3 Ligand/FLT3L (E. coli) 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.
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).
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What are the differences between Recombinant Human Flt-3 Ligand/FLT3L Protein (Catalog # 308-FK), Recombinant Human Flt-3 Ligand/FLT3L Protein (Catalog # 308-FKN), and Recombinant Human Flt-3 Ligand/FLT3L Protein, CF (Catalog # 308-FKE)?
Recombinant Human Flt-3 Ligand/FLT3L Protein (Catalog # 308-FK) and Recombinant Human Flt-3 Ligand/FLT3L Protein (Catalog # 308-FKN) share the same protein sequence, Thr27-Pro185 of Accession # AAA17999.1 but Catalog # 308-FK was expressed in Sf 21 baculovirus cells and Catalog # 308-FKN was produced in mammalian NS0 cells. Both of these proteins are glycosylated and available bottled with BSA or without BSA (Carrier-Free). Recombinant Human Flt-3 Ligand/FLT3L Protein, CF (Catalog # 308-FKE) was produced in an E. Coli expression system and has a sequence of (Thr27-Ala181, Accession # P49771.1), but may also contain an N-terminal Methionine followed by the expected sequence. In addition to being slightly shorter compared to 308-FKN and 308-FK, there is one difference in amino acid at position 72. Accession # AAA17999.1 has an Alanine at position 72, while Accession # P49771.1 has a Glycine in that position. All three versions of Recombinant Human Flt-3 Ligand/FLT3L proteins are routinely tested and have the same activity range in our QC testing assay: 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. R&D Systems also offers two GMP versions of this protein, an E. coli-derived animal-free version, Recombinant Human Flt-3 Ligand/FLT3L GMP Protein, CF (Catalog # 308E-GMP), and an Sf-21 baculovirus-derived version, Recombinant Human Flt-3 Ligand GMP Protein, CF (Catalog # 308-GMP). For information on the GMP proteins, we recommend consulting the product-specific pages for each protein. To determine which of these three proteins is most suitable for an application, we would recommend checking the Citations tab on the product-specific page.
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