Functional Assay
Activation of a human ST2-dependent NF-kappaB pathway. HEK-293 cells were transiently co-transfected with 500 ng of human ST2-containing vector (or empty vector as a negative control), a vector containing NF-kappaB-driven firefly luciferase, and a vector containing Renilla luciferase as a internal control in a 12-well plate. 40 hours after transfection, cells were treated with IL33 (human) (rec.) (His) at the concentration as indicated for 8 hours, followed by measurement with a dual luciferase assay.
Functional Assay
Pull down assay: a) pull down of hIL-33-His by hST2-Fc b) pull down of hST2-Fc by hIL-33-His 5 ug of hST2-Fc (or control Fc protein), 2 ug of hIL-33-His (or control His protein), and protein G resin (or anti-His resin) were incubated in 0.5ml RIPA buffer overnight at 4 degrees C. The precipitates were separated by SDS-PAGE, electro-transferred onto NC membrane, and immunoblotted for the presence of hIL-33-His or hST2-Fc with anti-His HRP or anti-hIgG HRP, respectively.
Functional Assay
Activation of a human ST2-dependent NF-kappaB pathway. HEK-293 cells were transiently co-transfected with 500 ng of human ST2-containing vector (or empty vector as a negative control), a vector containing NF-kappaB-driven firefly luciferase, and a vector containing Renilla luciferase as a internal control in a 12-well plate. 40 hours after transfection, cells were treated with IL33 (human) (rec.) (His) at the concentration as indicated for 8 hours, followed by measurement with a dual luciferase assay.
Functional Assay
Pull down assay: a) pull down of hIL-33-His by hST2-Fc b) pull down of hST2-Fc by hIL-33-His 5 ug of hST2-Fc (or control Fc protein), 2 ug of hIL-33-His (or control His protein), and protein G resin (or anti-His resin) were incubated in 0.5ml RIPA buffer overnight at 4 degrees C. The precipitates were separated by SDS-PAGE, electro-transferred onto NC membrane, and immunoblotted for the presence of hIL-33-His or hST2-Fc with anti-His HRP or anti-hIgG HRP, respectively.
Functional Assay
Activation of a human ST2-dependent NF-kappaB pathway. HEK-293 cells were transiently co-transfected with 500 ng of human ST2-containing vector (or empty vector as a negative control), a vector containing NF-kappaB-driven firefly luciferase, and a vector containing Renilla luciferase as a internal control in a 12-well plate. 40 hours after transfection, cells were treated with IL33 (human) (rec.) (His) at the concentration as indicated for 8 hours, followed by measurement with a dual luciferase assay.
Functional Assay
Pull down assay: a) pull down of hIL-33-His by hST2-Fc b) pull down of hST2-Fc by hIL-33-His 5 ug of hST2-Fc (or control Fc protein), 2 ug of hIL-33-His (or control His protein), and protein G resin (or anti-His resin) were incubated in 0.5ml RIPA buffer overnight at 4 degrees C. The precipitates were separated by SDS-PAGE, electro-transferred onto NC membrane, and immunoblotted for the presence of hIL-33-His or hST2-Fc with anti-His HRP or anti-hIgG HRP, respectively.
Functional Assay
Activation of a human ST2-dependent NF-kappaB pathway. HEK-293 cells were transiently co-transfected with 500 ng of human ST2-containing vector (or empty vector as a negative control), a vector containing NF-kappaB-driven firefly luciferase, and a vector containing Renilla luciferase as a internal control in a 12-well plate. 40 hours after transfection, cells were treated with IL33 (human) (rec.) (His) at the concentration as indicated for 8 hours, followed by measurement with a dual luciferase assay.
Functional Assay
Pull down assay: a) pull down of hIL-33-His by hST2-Fc b) pull down of hST2-Fc by hIL-33-His 5 ug of hST2-Fc (or control Fc protein), 2 ug of hIL-33-His (or control His protein), and protein G resin (or anti-His resin) were incubated in 0.5ml RIPA buffer overnight at 4 degrees C. The precipitates were separated by SDS-PAGE, electro-transferred onto NC membrane, and immunoblotted for the presence of hIL-33-His or hST2-Fc with anti-His HRP or anti-hIgG HRP, respectively.