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AdipoGen Life Sciences
DLL4 (human):Fc (human) (rec.)
290
CHF
CHF 290.00
In stock
AG-40A-0077Y-C01010 µgCHF 290.00
AG-40A-0077Y-C05050 µgCHF 530.00
Replaces AG-40A-0077
Figure 1: 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.
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.
Figure 2: 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-0077Y) (5μg/ml) or mCD137-Fc (5μg/ml) in PBS for 2 hours at 37°C. Plates were then used to differentiate 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-0077Y) (5μg/ml) or mCD137-Fc (5μg/ml) in PBS for 2 hours at 37°C. Plates were then used to differentiate MSCs.
Figure 3: 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-0077Y). Cells were stained with anti-human IgG(Fc specific) FITC conjugate for DLL4-Fc binding.
Figure 4: Adipogenesis inhibition of 3T3L-1 cells.
Figure 5: Induction of Hes-1 with the treatment of recombinant human DLL4-Fc (Prod. No. AG-40A-0077Y).
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.
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.
Figure 6: 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.
| Product Details | |
|---|---|
| Synonyms | Delta-like Protein 4; Delta4 |
| Product Type | Protein |
| 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 | ~100kDa (SDS-PAGE) |
| Purity | ≥95% (SDS-PAGE) |
| Endotoxin Content | <0.01EU/μg purified protein (LAL test; Lonza). |
| Concentration |
After reconstitution: for 10µg size: 0.1mg/ml for 50µg size: 1 mg/ml. |
| Reconstitution |
10µg size: Reconstitute with 100µl sterile endotoxin-free water. 50µg size: Reconstitute with 50µl sterile endotoxin-free water. |
| Formulation | Lyophilized. Contains PBS + 0.5 % Trehalose. |
| Protein Negative Control | |
| Other Product Data |
UniProt link Q9NR61: DLL4 (human) [Precursor] |
| 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. Centrifuge lyophilized vial before opening and reconstitution. 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. Working aliquots are stable for up to 3 months when stored at -20°C. |
| Documents | |
| MSDS |
 Download PDF |
| Product Specification Sheet | |
| Datasheet |
Download PDF |
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
- 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)
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