Anti-TRIF (TICAM1) 抗体 [EPR24995-152] (BSA and Azide free) (ab302563)

Western blot: Anti-TICAM1 antibody [EPR24995-152] (ab302562) staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab302562 was shown to bind specifically to TICAM1. A band was observed at 85 kDa in wild-type U-87 MG cell lysates with no signal observed at this size in TICAM1 knockout cell line. To generate this image, wild-type and TICAM1 knockout U-87 MG cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5% milk in TBS-0.1% Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

This data was developed using ab302562, the same antibody clone in a different buffer formulation.

This data was developed using ab302562, the same antibody clone in a different buffer formulation.

Diluting and blocking buffer and concentration: 5% NFDM/TBST. 

Lysates were freshly made and used for Western Blotting immediately to minimize protein degradation.
Negative control: HL-60.

This data was developed using ab302562, the same antibody clone in a different buffer formulation.

Diluting and blocking buffer and concentration: 5% NFDM/TBST. 

Lysates were freshly made and used for Western Blotting immediately to minimize protein degradation.

The identity of the lower MW band at approximately 18 kDa is unknown.

This data was developed using ab302562, the same antibody clone in a different buffer formulation.

TRIF (TICAM1) was immunoprecipitated from 0.35 mg Raji (human Burkitt's lymphoma b lymphocyte) whole cell lysate with ab302562 at 1/30 dilution (2 µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab302562 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1: Raji (human burkitt's lymphoma b lymphocyte) whole cell lysate 10 µg (Input)

Lane 2: ab302562 IP in Raji whole cell lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab302562 in Raji whole cell lysate

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 3 minutes

 Observed M_W (kDa): 98.