DFNA5 / GSDME Recombinant Rabbit Monoclonal Antibody [PSH09-51] - BSA and Azide free

☑ Cell treatment (CT),Relative expression (RE)

Western blot analysis of DFNA5 / GSDME on different lysates with Rabbit anti-DFNA5 / GSDME antibody (HA751298) at 1/2,000 dilution.

Lane 1: SH-SY5Y cell lysate
Lane 2: SH-SY5Y treated with 4μg/mL Doxorubicin for 24 hours cell lysate
Lane 3: SW480 cell lysate (negative)

Lysates/proteins at 20 µg/Lane.

Predicted band size: 55 kDa
Observed band size: 55/25 kDa

Exposure time: 3 minutes; ECL: K1801;

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 (HA751298) at 1/2,000 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.

☑ Cell treatment (CT)

Western blot analysis of DFNA5 / GSDME on different lysates with Rabbit anti-DFNA5 / GSDME antibody (HA751298) at 1/2,000 dilution.

Lane 1: 293T transfected with empty control cell lysate
Lane 2: 293T transfected with mouse GSDME (C-terminal) cell lysate
Lane 3: 293T transfected with mouse GSDME (N-terminal) cell lysate

Lysates/proteins at 10 µg/Lane.

Exposure time: 51 seconds; ECL: K1801;

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 (HA751298) at 1/2,000 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.

☑ Cell treatment (CT)

DFNA5 / GSDME was immunoprecipitated from 0.2 mg SH-SY5Y treated with 4μg/mL Doxorubicin for 24 hours cell lysate with HA751298 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA751298 at 1/1,000 dilution. Mouse Anti-Rabbit IgG kappa light chain secondary antibody (M1208-2) at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: SH-SY5Y treated with 4μg/mL Doxorubicin for 24 hours cell lysate (input)
Lane 2: HA751298 IP in SH-SY5Y treated with 4μg/mL Doxorubicin for 24 hours cell lysate
Lane 3: Rabbit IgG instead of HA751298 in SH-SY5Y treated with 4μg/mL Doxorubicin for 24 hours cell lysate

Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 10 seconds; ECL: K1801