Anti-C3AR1 antibody [EPR31126-546]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-C3AR1 antibody [EPR31126-546] (AB327652)

Immunohistochemical analysis of paraffin-embedded (A) Wild type C57BL/6J mouse liver and (B) C3ar1 knockout mouse liver tissue labeling with ab327652 at 1/1000 (0.507 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on (A) Wild type C57BL/6J mouse liver, no staining on (B) C3ar1 knockout mouse 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

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-C3AR1 antibody [EPR31126-546] (AB327652)

Immunohistochemical analysis of paraffin-embedded Mouse breast cancer tissue labeling with ab327652 at 1/1000 (0.507 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on immune cells of mouse breast 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-C3AR1 antibody [EPR31126-546] (AB327652)

Immunohistochemical analysis of paraffin-embedded Mouse skeletal muscle tissue labeling with ab327652 at 1/1000 (0.507 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : Positive staining on scattered immune cells of 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

• Flow Cyt

Lab

Flow Cytometry - Anti-C3AR1 antibody [EPR31126-546] (AB327652)

Flow cytometric analysis of Mouse bone marrow culture overnight labelling with ab327652 at 1/50 dilution (1ug) / Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/5000 dilution was used as the secondary antibody.

Gated on viable cells.
Cells were co-stained with CD3 conjugated to Alexa Fluor®488.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-C3AR1 antibody [EPR31126-546] (AB327652)

Immunohistochemical analysis of paraffin-embedded (A) HEK-293T (human embryonic kidney epithelial cell) transfected with a C3AR1 expression vector containing a his tag and (B) HEK-293T cells transfected with empty vector containing a myc-His-tag tissue labeling with ab327652 at 1/1000 (0.507 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on (A) HEK-293T transfected with a C3AR1 expression vector containing a his tag, no staining on (B) HEK-293T cells transfected with empty vector containing a myc-His-tag.

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-C3AR1 antibody [EPR31126-546] (AB327652)

Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labeling with ab327652 at 1/1000 (0.507 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on immune cells of mouse colon.

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

Lab

Flow Cytometry - Anti-C3AR1 antibody [EPR31126-546] (AB327652)

Flow cytometric analysis of Mouse bone marrow culture overnight labelling with ab327652 at 1/50 dilution (1ug) / Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/5000 dilution was used as the secondary antibody.

Gated on viable cells.
Cells were co-stained with CD14 conjugated to Brilliant Violet 421.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-C3AR1 antibody [EPR31126-546] (AB327652)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized J774A.1 (mouse reticulum cell sarcoma monocyte/macrophage) and J774A.1 treated with Lipopolysaccharides (1ug/ml) overnight cells labelling with ab327652 at 1/500 (9.68 μg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing increased membranous staining in J774A.1 treated with Lipopolysaccharides (1ug/ml) overnight (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab7291 Anti-alpha Tubulin mouse monoclonal antibody was used to counterstain tubulin at 1/1000 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-C3AR1 antibody [EPR31126-546] (AB327652)

Immunohistochemical analysis of paraffin-embedded Mouse Mantle cell lymphoma tissue labeling with ab327652 at 1/1000 (0.507 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on immune cells of mouse Mantle 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

Lab

Flow Cytometry - Anti-C3AR1 antibody [EPR31126-546] (AB327652)

Flow cytometric analysis of J774A.1 (mouse reticulum cell sarcoma monocyte/macrophage) labelling with ab327652 at 1/50 dilution (1ug) / 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.

Gated on viable cells.