ProDots Recombinant Human Flt-3 Ligand/FLT3L Protein
ProDots Recombinant Human Flt-3 Ligand/FLT3L Protein Summary
Pre-aliquoted, lyophilized protein dots for use in cell culture mediumFeatures and Benefits
- Rolls out of bottle for easy cell culture medium preparation
- Eliminates time spent aliquoting
- One protein dot can be used to make 500 mL of 10 ng/mL cell culture medium
ProDots® proteins are R&D Systems industry-leading quality, highly bioactive proteins packaged into easy-to-use lyophilized “dots”. Designed to be stable at 4 ºC and to dissolve instantly, ProDots® proteins can be rolled into cell culture media to quickly produce media with a desired working protein concentration.
ProDots® proteins are designed to reduce cell culture media preparation time while ensuring optimal protein recovery and bioactivity. Unlike standard lyophilized protein, which is shipped as a diffuse residue within a vial, ProDots® proteins are neatly bundled into the shape of a ball. In this form ProDots® proteins can be rolled directly into cell culture media. ProDots® proteins rehydrate instantly in media to maximize the recovery of purchased protein and to reliably reconstitute it at a desired concentration for use in cell culture media. ProDots® proteins can be conveniently stored at room temperature where they will remain stable for 6 months or at 4 ºC where they will remain stable for 12 months. The accessible storage and high stability of ProDots® proteins makes them ideal for quick and easy preparation of cell culture media.
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.
|Reconstitution||For a stock solution, reconstitute at 100 μg/mL in sterile PBS, or simply roll ProDot® directly into cell culture medium for immediate use.
|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.
• 6 months from date of receipt at room temperature as supplied.
• 12 months from date of receipt at 2-8 °C as supplied.
• 1 month at 2-8 °C under sterile conditions after reconstitution.
• 3 months at -20 to -80 °C under sterile conditions after reconstitution.
ProDots®Recombinant Human Flt-3 Ligand 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.
1 μg/lane of ProDots®Recombinant Human Flt-3 Ligand was resolved with SDS-PAGE under reducing (R) conditions and visualized with silver staining, showing major bands at 20-24 kDa. The multiple bands are due to variable glycosylation of the protein.
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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