Functional Assay
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 1 uM Dexamethasone, 0.5mM IBMX, 10 ug/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 ug/ml) dissolved in DPBS was added to the differentiation medium. These plates were then used to differentiate 3T3L-1 Cells.
Functional Assay
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 1 uM Dexamethasone, 0.5mM IBMX, 10 ug/m lnsulin, 100 uM 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 (5 ug/ml) or mCD137-Fc (5 ug/ml) in PBS for 2 hours at 37 degrees C. Plates were then used to differentiate MSCs.
Functional Assay
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 ug/ml of human GITR-Fc or human DLL4-Fc FITC conjugate for DLL4-Fc binding.
Functional Assay
Adipogenesis inhibition of 3T3L-1 cells.
Functional Assay
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 5 ug/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.
Functional Assay
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 1 uM Dexamethasone, 0.5mM IBMX, 10 ug/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 ug/ml) dissolved in DPBS was added to the differentiation medium. These plates were then used to differentiate 3T3L-1 Cells.
Functional Assay
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 1 uM Dexamethasone, 0.5mM IBMX, 10 ug/m lnsulin, 100 uM 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 (5 ug/ml) or mCD137-Fc (5 ug/ml) in PBS for 2 hours at 37 degrees C. Plates were then used to differentiate MSCs.
Functional Assay
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 ug/ml of human GITR-Fc or human DLL4-Fc FITC conjugate for DLL4-Fc binding.
Functional Assay
Adipogenesis inhibition of 3T3L-1 cells.
Functional Assay
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 5 ug/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.
Functional Assay
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 1 uM Dexamethasone, 0.5mM IBMX, 10 ug/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 ug/ml) dissolved in DPBS was added to the differentiation medium. These plates were then used to differentiate 3T3L-1 Cells.
Functional Assay
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 1 uM Dexamethasone, 0.5mM IBMX, 10 ug/m lnsulin, 100 uM 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 (5 ug/ml) or mCD137-Fc (5 ug/ml) in PBS for 2 hours at 37 degrees C. Plates were then used to differentiate MSCs.
Functional Assay
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 ug/ml of human GITR-Fc or human DLL4-Fc FITC conjugate for DLL4-Fc binding.
Functional Assay
Adipogenesis inhibition of 3T3L-1 cells.
Functional Assay
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 5 ug/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.
Functional Assay
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 1 uM Dexamethasone, 0.5mM IBMX, 10 ug/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 ug/ml) dissolved in DPBS was added to the differentiation medium. These plates were then used to differentiate 3T3L-1 Cells.
Functional Assay
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 1 uM Dexamethasone, 0.5mM IBMX, 10 ug/m lnsulin, 100 uM 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 (5 ug/ml) or mCD137-Fc (5 ug/ml) in PBS for 2 hours at 37 degrees C. Plates were then used to differentiate MSCs.
Functional Assay
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 ug/ml of human GITR-Fc or human DLL4-Fc FITC conjugate for DLL4-Fc binding.
Functional Assay
Adipogenesis inhibition of 3T3L-1 cells.
Functional Assay
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 5 ug/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.