Anti-TLR2 antibody [EPR30565-520]

• mIHC

Lab

Multiplex immunohistochemistry - Anti-TLR2 antibody [EPR30565-520] (AB325499)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human liver tissue staining TLR2 with ab325499 at a 1 : 100 (5.0 µg/ml) dilution, ab183322 anti-CD14 used at 1 : 100 (1.22 µg/ml) dilution and ab245118 anti-CD8 alpha used at a 1 : 500 (0.75 µg/ml) dilution.

Panel A : anti-TLR2 (green; Opal™520), anti-CD14 (magenta; Opal™570) and anti-CD8 alpha (yellow; Opal™690) on human liver.
Panel B : anti-TLPR2 staining immune cells in human liver.
Panel C : anti-CD14 staining macrophages in human liver.
Panel D : anti-CD8 alpha staining T lymphocytes in human liver.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab325499, ab183322 and ab245118 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Nuclear counter stain with DAPI.

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-TLR2 antibody [EPR30565-520] (AB325499)

Immunohistochemical analysis of paraffin-embedded Human colon cancer tissue labeling TLR2 with ab325499 at 1/100 (5.0 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in human colon cancer (PMID : 30532554).

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-TLR2 antibody [EPR30565-520] (AB325499)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling TLR2 with ab325499 at 1/100 (5.0 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in Kupffer cells at human liver.

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

• WB

Lab

Western blot - Anti-TLR2 antibody [EPR30565-520] (AB325499)

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

The identity of the bands lower than 75 kDa are unknown.

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

All lanes:

Western blot - Anti-TLR2 antibody [EPR30565-520] (ab325499) at 1/1000 dilution

Lane 1:

Human lung tissue lysate at 20 µg

Lane 2:

Human liver 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: 89 kDa,36 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-TLR2 antibody [EPR30565-520] (AB325499)

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

Negative control : HeLa (PMID : 12819075), PANC-1 (PMID : 36232715), 293T.

Low expression : Caco-2 (PMID : 38170495).

The identity of the bands higher than 150 kDa and lower than 60 kDa are unknown.

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

Exposure time : Lanes 1-3 : 180 seconds; Lane 4 : 70 seconds; Lanes 5-7 : 180 seconds.

All lanes:

Western blot - Anti-TLR2 antibody [EPR30565-520] (ab325499) at 1/1000 dilution

Lane 1:

DLD-1 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

Caco-2 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

THP-1 (human monocytic leukemia monocyte) whole cell lysate at 20 µg

Lane 5:

BxPC-3 (human pancreas adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 6:

293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 7:

PANC-1 (human pancreatic epithelioid carcinoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

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

Observed band size: 89 kDa,36 kDa

false

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TLR2 antibody [EPR30565-520] (AB325499)

Immunohistochemical analysis of paraffin-embedded Human cardiac muscle tissue labeling TLR2 with ab325499 at 1/100 (5.0 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : No staining in human cardiac 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.

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