重组Anti-C3a R抗体[EPR28765-1] (ab317321)

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling C3a R with ab317321 at 1/1000 (0.517 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on microglia of human cerebrum (PMID: 28002390).
The section was incubated with ab317321 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

Immunohistochemical analysis of paraffin-embedded Human astrocytoma tissue labeling C3a R with ab317321 at 1/1000 (0.517 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on microglia of human astrocytoma (PMID: 37174113).
The section was incubated with ab317321 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

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling C3a R with ab317321 at 1/1000 (0.517 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on kupffer cells of human liver (PMID: 24974766).
The section was incubated with ab317321 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

Immunohistochemical analysis of paraffin-embedded Human placenta tissue labeling C3a R with ab317321 at 1/1000 (0.517 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on hofbauer cells of human placenta (PMID 22901903).
The section was incubated with ab317321 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

Immunohistochemical analysis of paraffin-embedded (A) HEK-293T (human embryonic kidney epithelial cell) transfected with a C3AR1 expression vector containing a his tag. (B) HEK-293T transfected with empty vector containing a his tag. tissue labeling C3a R with ab317321 at 1/1000 (0.517 ug/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 transfected with empty vector containing a his tag.
The section was incubated with ab317321 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

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human tonsil tissue staining C3a R with ab317321 at a 1:1000 (0.52 ug/ml) dilution, ab183322 anti-CD14 used at 1:100 (1.22 ug/ml) dilution and ab16669 anti-CD3 used at a 1:150 (0.06 ug/ml) dilution.

Panel A: merged staining of anti-C3AR1 (green; Opal™520), anti-CD14 (red; Opal™570), anti-CD3 (magenta; Opal™690) on human tonsil.
Panel B: anti-C3AR1 stained on immune cells.
Panel C: anti-CD14 stained on macrophages.
Panel D: anti-CD3 stained on T lymphocytes.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining: in the order of ab317321, ab183322 and ab16669 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.