Anti-TLR9 antibody [EPR14964-2]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TLR9 antibody [EPR14964-2] (AB187148)

Immunohistochemistry analysis of paraffin-embedded Human breast carcinoma tissue sections labelling TLR9 with ab187148 at 1/1000 dilution. The section was incubated with ab187148 for 30 mins at room temperature. Ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used as the secondary antibody. Sections were counterstained with Hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins. Positive staining on some immune stroma cells in human breast carcinoma tissue. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TLR9 antibody [EPR14964-2] (AB187148)

Immunohistochemistry analysis of paraffin-embedded Human tonsil tissue sections labelling TLR9 with ab187148 at 1/1000 dilution. The section was incubated with ab187148 for 30 mins at room temperature. Ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used as the secondary antibody. Sections were counterstained with Hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins. Positive staining on some immune cells in human tonsil tissue. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

• WB

Supplier Data

Western blot - Anti-TLR9 antibody [EPR14964-2] (AB187148)

Blocking and dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-TLR9 antibody [EPR14964-2] (ab187148) at 1/5000 dilution

Lane 1:

Daudi cell lysate at 10 µg

Lane 2:

Raji cell lysate at 10 µg

Lane 3:

Ramos cell lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 115 kDa

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• WB

Lab

Western blot - Anti-TLR9 antibody [EPR14964-2] (AB187148)

False colour image of Western blot : Anti-TLR9 antibody [EPR14964-2] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab187148 was shown to bind specifically to TLR9. A band was observed at 140 kDa in wild-type Raji cell lysates with no signal observed at this size in TLR9 knockout cell line ab280879 (knockout cell lysate ab282939). To generate this image, wild-type and TLR9 knockout Raji 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-TLR9 antibody [EPR14964-2] (ab187148) at 1/1000 dilution

Lane 1:

Wild-type Raji cell lysate at 20 µg

Lane 2:

TLR9 knockout Raji cell lysate at 20 µg

Predicted band size: 115 kDa

Observed band size: 140 kDa

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