Anti-TIL/TLR1 antibody [EPR2075]

• Flow Cyt

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Flow Cytometry - Anti-TIL/TLR1 antibody [EPR2075] (AB68158)

Flow cytometric analysis of TIL/TLR1 in Daudi cells using ab68158 at 1/60 dilution (pink) or a rabbit IgG (green).

• IP

Lab

Immunoprecipitation - Anti-TIL/TLR1 antibody [EPR2075] (AB68158)

TIL/TLR1 was immunoprecipitated from 0.35 mg Ramos (Human Burkitt's lymphoma B lymphocyte) whole cell lysate 10 μg with ab68158 at 1/50 dilution (2μg). VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : Ramos (Human Burkitt's lymphoma B lymphocyte) whole cell lysate 10 μg

Lane 2 : ab68158 IP in Ramos whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab68158 in Ramos whole cell lysate

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

All lanes:

Immunoprecipitation - Anti-TIL/TLR1 antibody [EPR2075] (ab68158)

Predicted band size: 90 kDa

Observed band size: 90-125 kDa

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• WB

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Western blot - Anti-TIL/TLR1 antibody [EPR2075] (AB68158)

All lanes:

Western blot - Anti-TIL/TLR1 antibody [EPR2075] (ab68158) at 1/1000 dilution

Lane 1:

Daudi cell lysate at 10 µg

Lane 2:

Ramos cell lysate at 10 µg

Secondary

All lanes:

HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 90 kDa

Observed band size: 125 kDa,90 kDa

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• WB

Lab

Western blot - Anti-TIL/TLR1 antibody [EPR2075] (AB68158)

Western blot : Rabbit Monoclonal [EPR2075] to TIL/TLR1 ab68158 staining at 1/1000 dilution, shown in green; Mouse anti alpha Tubulin (ab7291) loading control staining at 1/20,000 dilution, shown in magenta.

A band was observed at 99 kDa in Wild-type A549 cell lysates with no signal observed at this size in TLR1 knockout A549 cell line.

To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc 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 800CW & Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-TIL/TLR1 antibody [EPR2075] (ab68158) at 1/1000 dilution

Lane 1:

Wild-type A549 at 20 µg

Lane 2:

TLR1 knockout A549 at 20 µg

Lane 2:

Western blot - Human TLR1 knockout A549 cell line (<a href='/en-us/products/cell-lines/human-tlr1-knockout-a549-cell-line-ab289019'>ab289019</a>) at 20 µg

Lane 3:

THP-1 at 20 µg

Lane 4:

HeLa Nuclear at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 90 kDa

Observed band size: 99 kDa

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