WB
Western blot analysis of Phospho-TAK1 (S412) on different lysates with Rabbit anti-Phospho-TAK1 (S412) antibody at 1/5,000 dilution.Lysates/proteins at 20 µg/Lane.Exposure time: 10 seconds; 4-20% SDS-PAGE gel.Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/5,000 dilution was used in primary antibody dilution at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
FC
Flow cytometric analysis of HeLa cells untreated (left) / treated with 100nM Calyculin A for 30 minutes (right) labeling Phospho-TAK1 (S412).Cells were fixed and permeabilized. Then stained with the primary antibody (1/2,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
FC
Flow cytometric analysis of NIH/3T3 cells untreated (left) / treated with 100nM Calyculin A for 30 minutes (right) labeling Phospho-TAK1 (S412).Cells were fixed and permeabilized. Then stained with the primary antibody (1/2,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).