Anti-TLR3 antibody [EPR27050-89]

• WB

Supplier Data

Western blot - Anti-TLR3 antibody [EPR27050-89] (AB307442)

Blocking and diluting buffer and concentration : 5% NFDM/TBSTThe molecular weight observed is consistent with what has been described in the literature (PMID : 28717003). Exposure time : 103 seconds

All lanes:

Western blot - Anti-TLR3 antibody [EPR27050-89] (ab307442) at 1/1000 dilution

Lane 1:

J774A.1 (mouse reticum cell sarcoma monocyte macrophage) whole cell lysate at 20 µg

Lane 2:

Untreated HT-29 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

HT-29 treated with 1µg/ml poly(I:C) overnight whole cell lysate at 20 µg

Lane 4:

Untreated HCT116 (human colorectal carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 5:

HCT116 treated with 1µg/ml poly(I:C) overnight whole cell lysate at 20 µg

Lane 6:

Human tonsil tissue lysate at 20 µg

Lane 7:

Mouse spleen tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 75 kDa,120 kDa,150 kDa

false

Exposure time: 103s

• WB

Lab

Western blot - Anti-TLR3 antibody [EPR27050-89] (AB307442)

Western blot : ab307442 staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, the antibody was shown to bind specifically to TLR3. A band was observed at 135 kDa in wild-type A549 cell lysates with no signal observed at this size in TLR3 knockout cell line ab288996. To generate this image, wild-type and TLR3 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution 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.

All lanes:

Western blot - Anti-TLR3 antibody [EPR27050-89] (ab307442) at 1/1000 dilution

Lane 1:

Wild-type A549 control for Poly (I:C) (0 ug/mL, 24 h) cell lysate at 20 µg

Lane 2:

Wild-type A549 Treated Poly (I:C) (1 ug/mL, 24 h) cell lysate at 20 µg

Lane 3:

Wild-type A549 control for IFNb (0 IU/mL, 6 h) cell lysate at 20 µg

Lane 4:

Wild-type A549 Treated IFNb (100 IU/mL, 6 h) cell lysate at 20 µg

Lane 5:

TLR3 knockout A549 control for Poly (I:C) (0 ug/mL, 24 h) cell lysate at 20 µg

Lane 6:

TLR3 knockout A549 Treated Poly (I:C) (1 ug/mL, 24 h) cell lysate at 20 µg

Lane 7:

TLR3 knockout A549 control for IFNb (0 IU/mL, 6 h) cell lysate at 20 µg

Lane 8:

TLR3 knockout A549 Treated IFNb (100 IU/mL, 6 h) cell lysate at 20 µg

Observed band size: 135 kDa

false