DLL4 (human):Fc (human) (rec.)
3T3L-1 Cells(mouse pre-adipocyte cells) were maintained in DMEM, supplemented with 10% fetal bovine serum and penicillin-streptomycin. For differentiation of 3T3L-1 Cells, 3T3L-1 Cells were cultured in adipogenic medium which was growth medium supplemented with 1μM Dexamethasone, 0.5mM IBMX, 10μg/ml lnsulin (day 0). Medium was changed every 2 days. Staining with Oil Red O was typically performed on day 7. Cells were washed twice with PBS, fixed with 3.7% formalin, and stained with 0.5% filtered Oil Red O in propylene glycol. For negative controls, mouse TNF-α (20ng/ml) was added. Recombinant human DLL4-Fc (5μg/ml) dissolved in DPBS was added to the differentiation medium. These plates were then used to differentiate 3T3L-1 Cells.
MSCs (Mesenchymal stem cells) were maintained in DMEM, supplemented with 10% fetal bovine serum, penicilin-streptomycin and glutamine. For differentiation of MSCs, MSCs were cultured in adipogenic medium which was growth medium supplemented with 1μM Dexamethasone, 0.5mM IBMX, 10μg/m lnsulin, 100μM Indomethacin (day 1). Medium was changed every 3 days. Staining with Oil Red O was typically performed on day 30. For negative controls, TNF-α (20ng/ml) was added. To immobilize Notch ligands on the plastic surface of the culture plates, plates were incubated with a solution of hDLL4-Fc (Prod. No. AG-40A-0077) (5μg/ml) or mCD137-Fc (5μg/ml) in PBS for 2 hours at 37°C. Plates were then used to differentiate MSCs.
A mouse preadpipocyte cell line, 3T3L1, was stimulated with 5μg/ml of human DLL4-Fc as in indicated time points and each cell lysate was prepared and subjected to western blot by using anti-mouse Hes1 or GAPDH.
|Synonyms||Delta-like Protein 4; Delta4|
|Source/Host||HEK 293 cells|
|Sequence||Signal peptide and extracellular domain of human DLL4 (aa 1-529) are fused at the C-terminus to the Fc portion of human IgG1.|
|Specificity||Interacts with human Notch1 (as confirmed by flow cytometry).|
|Biological Activity||Inhibits adipogenesis of 3T3L-1 cells and mesenchymal stem cells (MSCs). Induces the Notch target gene HES-1 when coated on a plate at 1µg/ml.|
|Endotoxin Content||<0.01EU/μg purified protein (LAL test; Lonza).|
10µg size: 0.1mg/ml after reconstitution.
50µg size: 1mg/ml after reconstitution.
10µg size: Reconstitute with 100µl sterile water.
50µg size: Reconstitute with 50µl sterile water.
|Formulation||Lyophilized. Contains PBS + 0.5 % Trehalose.|
|Other Product Data||UniProt link Q9NR61: DLL4 (human) [Precursor]|
|Shipping and Handling|
|Short Term Storage||+4°C|
|Long Term Storage||-20°C|
After reconstitution, prepare aliquots and store at -20°C.
Avoid freeze/thaw cycles.
PBS containing at least 0.1% BSA should be used for further dilutions.
|Use/Stability||Stable for at least 6 months after receipt when stored at -20°C.|
|Product Specification Sheet|
The Notch ligand delta-like protein 4 (DLL4) is expressed highly and selectively within the arterial endothelium and has been shown to function as a ligand for Notch1 and Notch4. It is induced by VEGF as a negative feedback regulator and acts to prevent overexuberant angiogenic sprouting, promoting the timely formation of a well differentiated vascular network. DLL4-Notch1 signaling regulates the formation of appropriate numbers of tip cells to control vessel sprouting and branching in the mouse retina.
- Jagged2 acts as a Delta-like Notch ligand during early hematopoietic cell fate decisions: I. Van de Walle, et al.; Blood 117, 4449 (2011)
- Notch regulates BMP responsiveness and lateral branching in vessel networks via SMAD6: K.P. Mouillesseaux, et al.; Nat. Commun. 7, 13247 (2016)
- Notch ligands regulate the muscle stem-like state ex vivo but are not sufficient for retaining regenerative capacity: H. Sakai, et al.; PLoS ONE 12, e0177516 (2017)