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DLL4 (human):Fc (human) (rec.)

Replaces AG-40A-0077
AG-40A-0077Y-C010 10 µg INQ
AG-40A-0077Y-C050 50 µg INQ
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Additional Information

Product Data
Synonyms Delta-like Protein 4; Delta4
Properties
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.
Crossreactivity Human
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.
MW ~80kDa (SDS-PAGE)
Purity ≥95% (SDS-PAGE)
Endotoxin Content <0.01EU/μg purified protein (LAL test; Lonza).
Concentration 10µg size: 0.1mg/ml after reconstitution.
50µg size: 1mg/ml after reconstitution.
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]
Product Type Protein
Shipping and Handling
Shipping BLUE ICE
Short Term Storage +4°C
Long Term Storage -20°C
Handling Advice 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.
Documents
MSDS Download Document Download

Product Description

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.

Product References

  1. Jagged2 acts as a Delta-like Notch ligand during early hematopoietic cell fate decisions: I. Van de Walle, et al.; Blood 117, 4449 (2011)
Adipogenesis inhibition of 3T3L-1 cells.<br /> 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 1&mu;M Dexamethasone, 0.5mM IBMX, 10&mu;g/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-&alpha; (20ng/ml) was added. Recombinant human DLL4-Fc (5&mu;g/ml) dissolved in DPBS was added to the differentiation medium. These plates were then used to differentiate 3T3L-1 Cells.
Adipogenesis inhibition of 3T3L-1 cells.
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.
Adipogenesis inhibition of MSCs.<br />
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 1&mu;M Dexamethasone, 0.5mM IBMX, 10&mu;g/m lnsulin, 100&mu;M Indomethacin (day 1). Medium was changed every 3 days. Staining with Oil Red O was typically performed on day 30. For negative controls, TNF-&alpha; (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) (5&mu;g/ml) or mCD137-Fc (5&mu;g/ml) in PBS for 2 hours at 37°C. Plates were then used to differentiate MSCs.
Adipogenesis inhibition of MSCs.
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.
Interaction of human Notch1 with human DLL4. HEK293 cells transfected with a human Notch1 or a human GITR ligand expressing vector were incubated with 25&mu;g/ml of human GITR-Fc or human DLL4-Fc (Prod. No. AG-40A-0077). Cells were stained with anti-human IgG(Fc specific) FITC conjugate for DLL4-Fc binding.
Interaction of human Notch1 with human DLL4. HEK293 cells transfected with a human Notch1 or a human GITR ligand expressing vector were incubated with 25μg/ml of human GITR-Fc or human DLL4-Fc (Prod. No. AG-40A-0077). Cells were stained with anti-human IgG(Fc specific) FITC conjugate for DLL4-Fc binding.
Adipogenesis inhibition of 3T3L-1 cells.
Adipogenesis inhibition of 3T3L-1 cells.
Induction of Hes-1 with the treatment of recombinant human DLL4-Fc (Prod. No. AG-40A-0077).<br /> A mouse preadpipocyte cell line, 3T3L1, was stimulated with 5&mu;g/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.
Induction of Hes-1 with the treatment of recombinant human DLL4-Fc (Prod. No. AG-40A-0077).
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.
50&mu;g of cell lysates derived from hDLL4-Fc or non-treated 3T3L1 cells, which had been either differentiated or undifferentiated, were subjected to western blot by using a mouse adiponectin antibody.
50μg of cell lysates derived from hDLL4-Fc or non-treated 3T3L1 cells, which had been either differentiated or undifferentiated, were subjected to western blot by using a mouse adiponectin antibody.
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