Phospho-STING (S366) Recombinant Rabbit Monoclonal Antibody [PSH09-72] - BSA and Azide free

☑ Cell treatment (CT)

Western blot analysis of Phospho-STING (S366) on different lysates with Rabbit anti-Phospho-STING (S366) antibody (HA751314) at 1/2,000 dilution.

Lane 1: THP-1 treated with 80nM TPA for 16 hours cell lysate
Lane 2: THP-1 treated with 80nM TPA for 16 hours then transfected with 5μg/mL Poly (dA:dT) for 3 hours cell lysate
Lane 3: THP-1 treated with 80nM TPA for 16 hours then transfected with 5μg/mL Poly (dA:dT) for 3 hours cell lysate, then the membrane treated with λpp for 1 hour

Lysates/proteins at 20 µg/Lane.

Predicted band size: 42 kDa
Observed band size: 42 kDa

Exposure time: 59 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 (HA751314) 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.

Dot blot analysis of Phospho-STING (S366) on different peptides with Rabbit anti-Phospho-STING (S366) antibody (HA751314) at 1/2,000 dilution. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution for 1 hour at room temperature.

Lane 1: Unmodified STING peptide (negative)
Lane 2: Phospho-STING (S366) peptide (positive)

Proteins loading: 100ng, 50ng, 10ng, 5ng;

Blocking and dilution buffer: 5% NFDM/TBST;

Exposure time: 1 minute; ECL: K1801.

☑ Cell treatment (CT)

Phospho-STING (S366) was immunoprecipitated from 0.2 mg THP-1 treated with 80nM TPA for 16 hours then transfected with 5μg/mL Poly (dA:dT) for 3 hours cell lysate with HA751314 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA751314 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: THP-1 treated with 80nM TPA for 16 hours then transfected with 5μg/mL Poly (dA:dT) for 3 hours cell lysate (input)
Lane 2: HA751314 IP in THP-1 treated with 80nM TPA for 16 hours then transfected with 5μg/mL Poly (dA:dT) for 3 hours cell lysate
Lane 3: Rabbit IgG instead of HA751314 in THP-1 treated with 80nM TPA for 16 hours then transfected with 5μg/mL Poly (dA:dT) for 3 hours cell lysate

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