Anti-PAR1/Thrombin Receptor antibody [EPR28398-8]

• mIHC

Lab

Multiplex immunohistochemistry - Anti-PAR1/Thrombin Receptor antibody [EPR28398-8] (AB322457)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse liver tissue staining ALDH1L1 with ab307696 at a 1/5000 dilution, ab322457 anti-PAR1 used at 1/500 dilution and ab315474 anti-ABCB11 used at a 1/2000 dilution.

Panel A : merged staining of anti-ALDH1L1 (green; Opal^™520), anti-PAR1 (magenta; Opal^™690) and anti-ABCB11 (gray; Opal^™570) on mouse liver.
Panel B : anti-ALDH1L1 staining hepatocytes in mouse liver.
Panel C : anti-PAR1 staining endothelium in mouse liver.
Panel D : anti-ABCB11 staining bile canaliculi in mouse liver.
Nuclear DNA was labeled with DAPI (shown in blue).
The section was incubated in three rounds of staining : in the order of ab307696, ab322457 and ab315474 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAR1/Thrombin Receptor antibody [EPR28398-8] (AB322457)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling PAR1/Thrombin Receptor with ab322457 at 1/500 (1.054 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse spleen.
The section was incubated with ab322457 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

Lab

Immunohistochemistry - Anti-PAR1/Thrombin Receptor antibody [EPR28398-8] (AB322457)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse kidney tissue staining PAR1/Thrombin Receptor with ab322457 at a 1/500 dilution, ab306558 anti-SGLT2 used at 1/5000 dilution and ab212197 anti-Renin used at a 1/4000 dilution.

Panel A : merged staining of anti-PAR1/Thrombin Receptor (green; Opal™520), anti-SGLT2 (magenta; Opal™690) and anti-Renin (gray; Opal™570) on mouse kidney.

Panel B : anti-PAR1/Thrombin Receptor staining endothelium in mouse kidney.

Panel C : anti-SGLT2 staining the brush border of proximal tubules in mouse kidney.

Panel D : anti-Renin staining juxtaglomerular cells in mouse kidney.

Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab322457, ab306558 and ab212197 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.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAR1/Thrombin Receptor antibody [EPR28398-8] (AB322457)

Immunohistochemical analysis of paraffin-embedded Mouse lung tissue labeling PAR1/Thrombin Receptor with ab322457 at 1/500 (1.054 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse lung (PMID : 12548614).
The section was incubated with ab322457 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

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAR1/Thrombin Receptor antibody [EPR28398-8] (AB322457)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling PAR1/Thrombin Receptor with ab322457 at 1/500 (1.054 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse liver.
The section was incubated with ab322457 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

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAR1/Thrombin Receptor antibody [EPR28398-8] (AB322457)

Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling PAR1/Thrombin Receptor with ab322457 at 1/500 (1.054 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse kidney.
The section was incubated with ab322457 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

Supplier Data

Western blot - Anti-PAR1/Thrombin Receptor antibody [EPR28398-8] (AB322457)

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

PAR1 contains different glycosylation sites, the multiple bands may be due to glycosylation (PMID : 20368337).

This blot was developed using a high-sensitivity ECL substrate, allowing for the detection of proteins in the mid-femtogram range.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-PAR1/Thrombin Receptor antibody [EPR28398-8] (ab322457) at 1/1000 dilution

Lane 1:

Neuro-2a (mouse neuroblastoma neuroblast) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

Neuro-2a transfected with siRNA specifically targeting PAR1 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: 47-75 kDa,36 kDa

true

Exposure time: 81s

• WB

Supplier Data

Western blot - Anti-PAR1/Thrombin Receptor antibody [EPR28398-8] (AB322457)

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

Low expression : RAW 264.7, HL-1.

PAR1 contains different glycosylation sites, the multiple bands may be due to glycosylation (PMID : 20368337).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-PAR1/Thrombin Receptor antibody [EPR28398-8] (ab322457) at 1/1000 dilution

Lane 1:

Mouse dorsal galion tissue lysate at 20 µg

Lane 2:

Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 20 µg

Lane 3:

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

Lane 4:

HL-1 (mouse atrial muscle cell) whole cell lysate at 20 µg

Lane 5:

C8-D1A (mouse cerebellum astrocyte) whole cell lysate at 20 µg

Lane 6:

SW10 (mouse neuronal schwann cell) whole cell lysate at 20 µg

Lane 7:

NIH/3T3 (mouse embryonic fibroblast) 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: 48-75 kDa,36 kDa

false

Exposure time: 136s