Anti-DARC antibody [EPR29719-512]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DARC antibody [EPR29719-512] (AB325127)

Immunohistochemical analysis of paraffin-embedded Mouse bone marrow tissue labeling DARC with ab325127 at 1/500 (0.988 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on erythrocytes and endothelium in mouse bone marrow (PMID : 28553950). 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

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DARC antibody [EPR29719-512] (AB325127)

Immunohistochemical analysis of paraffin-embedded Mouse cardiac muscle tissue labeling DARC with ab325127 at 1/500 (0.988 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression : weak staining in mouse 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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DARC antibody [EPR29719-512] (AB325127)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling DARC with ab325127 at 1/500 (0.988 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on endothelium in 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

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DARC antibody [EPR29719-512] (AB325127)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling DARC with ab325127 at 1/500 (0.988 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on erythrocytes in mouse spleen (PMID : 28553950). 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

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Flow Cytometry - Anti-DARC antibody [EPR29719-512] (AB325127)

Flow cytometric analysis of Mouse PBMC (mouse primary peripheral blood mononuclear cell) cells labelling DARC with ab325127 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® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Gated on viable cells.

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-DARC antibody [EPR29719-512] (AB325127)

Flow cytometric analysis of Mouse blood cells labelling DARC with ab325127 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® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Gated on viable cells.
Cells were co-stained with anti mouse TER-119 conjugated to Alexa Fluor®647.

• WB

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Western blot - Anti-DARC antibody [EPR29719-512] (AB325127)

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

Low expression : testis, skeletal muscle, heart.

High expression : bone marrow (PMID : 37006247, 26908826, 28553950), spleen (PMID : 7699323).

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 37153544).

The identity of the higher than 250 kDa (in lane 4) is unknown.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

All lanes:

Western blot - Anti-DARC antibody [EPR29719-512] (ab325127) at 1/1000 dilution

Lane 1:

Mouse bone marrow tissue at 20 µg

Lane 2:

Mouse spleen tissue lysate at 20 µg

Lane 3:

Mouse testis tissue at 20 µg

Lane 4:

Mouse skeletal muscle tissue lysate at 20 µg

Lane 5:

Mouse heart tissue lysate at 20 µg

Secondary

All lanes:

Western blot at 1/20000 dilution

Observed band size: 37-50 kDa,124 kDa

false

Exposure time: 37s