Gasdermin D (N terminal) Recombinant Rabbit Monoclonal Antibody [PSH10-87]

☑ Cell treatment (CT)

Western blot analysis of Gasdermin D (N terminal) on different lysates with Rabbit anti-Gasdermin D (N terminal) antibody (HA723254) at 1/2,000 dilution.

Lane 1: THP-1 cell lysate
Lane 2: THP-1 treated with 100nM TPA overnight then add 100ng/mL LPS for 7 hours then add 1μg/mL BFA for 3 hours cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 30 kDa
Observed band size: 35 kDa

Exposure time: 2 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 (HA723254) 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)

Gasdermin D (N terminal) was immunoprecipitated from 0.2 mg THP-1 treated with 80nM TPA for 16 hours then treated with 5μg/mL poly(dA:dT) for 3 hours cell lysate with HA723254 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA723254 at 1/1,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: THP-1 treated with 80nM TPA for 16 hours then treated with 5μg/mL poly(dA:dT) for 3 hours cell lysate (input)
Lane 2: HA723254 IP in THP-1 treated with 80nM TPA for 16 hours then treated with 5μg/mL poly(dA:dT) for 3 hours cell lysate
Lane 3: Rabbit IgG instead of HA723254 in THP-1 treated with 80nM TPA for 16 hours then treated with 5μg/mL poly(dA:dT) for 3 hours cell lysate

Blocking/Dilution buffer: primary antibody dilution (K1803)
Exposure time: 10econds; ECL: K1801