Western blot analysis of TRADD on different lysates with Rabbit anti-TRADD antibody (HA722000) at 1/1,000 dilution. Lane 1: HL-60 cell lysate (20 µg/Lane) Lane 2: HeLa cell lysate (20 µg/Lane) Lane 3: K-562 cell lysate (20 µg/Lane) Lane 4: Ramos cell lysate (20 µg/Lane) Lane 5: Jurkat cell lysate (20 µg/Lane) Lane 6: C2C12 cell lysate (20 µg/Lane) Lane 7: PC-12 cell lysate (20 µg/Lane) Predicted band size: 34 kDa Observed band size: 34 kDa Exposure time: 2 minutes 18 seconds; ECL: K1802; 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 (HA722000) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

Immunocytochemistry analysis of MCF7 cells labeling TRADD with Rabbit anti-TRADD antibody (HA722000) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-TRADD antibody (HA722000) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.