Anti-TLR7 antibody [EPR26195-250]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TLR7 antibody [EPR26195-250] (AB325746)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling TLR7 with ab325746 at 1/500 (0.976 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse spleen. The primary antibody was incubated for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TLR7 antibody [EPR26195-250] (AB325746)

Immunohistochemical analysis of paraffin-embedded Mouse lung cancer tissue labeling TLR7 with ab325746 at 1/500 (0.976 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on interstitial cells of mouse lung cancer. The primary antibody was incubated for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TLR7 antibody [EPR26195-250] (AB325746)

Immunohistochemical analysis of paraffin-embedded Mouse T-cell lymphoma tissue labeling TLR7 with ab325746 at 1/500 (0.976 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse T-cell lymphoma. The primary antibody was incubated for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-TLR7 antibody [EPR26195-250] (AB325746)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage, Right) and WEHI-231 (mouse B cell lymphoma B lymphocyte, Left) cells labelling TLR7 with ab325746 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Negative control : WEHI-231.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-TLR7 antibody [EPR26195-250] (AB325746)

Flow cytometric analysis of 4% paraformaldehyde fixed 0.1% Tween-20 permeabilized Mouse bone marrow cells labelling TLR7 with ab325746 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Cells were surface stained with anti-CD11c conjugated to PE/CY7 and anti-CD45R/B220 conjugated to BV421. Then fixed with 2% PFA for 10 min followed by intracellularly staining with rabbit IgG or our antibody.
Gated on CD11c+ and CD45R/B220+ shows upper right, CD11c- and CD45R/B220- shows lower right.

• WB

Lab

Western blot - Anti-TLR7 antibody [EPR26195-250] (AB325746)

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

Negative control : heart, WEHI-231(PMID : 12202403).

The molecular weight observed is consistent with what has been described in the literature (PMID : 25648980).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

Exposure time : Lanes 1-3 : 180 seconds; Lanes 4-5 : 59 seconds.

All lanes:

Western blot - Anti-TLR7 antibody [EPR26195-250] (ab325746) at 1/1000 dilution

Lane 1:

Mouse lymph node tissue lysate at 20 µg

Lane 2:

Mouse spleen tissue lysate at 20 µg

Lane 3:

Mouse heart tissue lysate at 20 µg

Lane 4:

RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

Lane 5:

WEHI-231 (mouse b cell lymphoma b lymphocyte ) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 60 kDa,140 kDa,36 kDa

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• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TLR7 antibody [EPR26195-250] (AB325746)

Immunohistochemical analysis of paraffin-embedded Mouse skeletal muscle tissue labeling TLR7 with ab325746 at 1/500 (0.976 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on mouse skeletal muscle. The primary antibody was incubated for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins