Anti-CD204 antibody [EPR28270-88] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD204 antibody [EPR28270-88] - BSA and Azide free (AB314228)

This data was developed using ab314227, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Mouse lung cancer tissue labeling CD204 with ab314227 at 1/5000 (0.103 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on immune cells in mouse lung cancer.The section was incubated with ab314227 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-CD204 antibody [EPR28270-88] - BSA and Azide free (AB314228)

This data was developed using ab314227, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Mouse neuroendocrine carcinoma tissue labeling CD204 with ab314227 at 1/5000 (0.103 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on immune cells in mouse neuroendocrine carcinoma.The section was incubated with ab314227 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-CD204 antibody [EPR28270-88] - BSA and Azide free (AB314228)

This data was developed using ab314227, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Mouse lung tissue labeling CD204 with ab314227 at 1/5000 (0.103 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on macrophages in mouse lung.The section was incubated with ab314227 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-CD204 antibody [EPR28270-88] - BSA and Azide free (AB314228)

This data was developed using ab314227, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded (A) RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) cell pellets; (B) Neuro-2a (mouse neuroblastoma neuroblast) cell pellets. tissue labeling CD204 with ab314227 at 1/2000 (0.257 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on (A) RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) cell pellets, no staining on (B) Neuro-2a (mouse neuroblastoma neuroblast) cell pellets.The section was incubated with ab314227 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-CD204 antibody [EPR28270-88] - BSA and Azide free (AB314228)

This data was developed using ab314227, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labeling CD204 with ab314227 at 1/5000 (0.103 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on immune cells in mouse colon.The section was incubated with ab314227 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

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-CD204 antibody [EPR28270-88] - BSA and Azide free (AB314228)

This data was developed using ab314227, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Mouse bone marrow cells labelling CD204 with ab314227 at 1/50 (10.26 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing positive staining in mouse bone marrow treated with lipopolysaccharide (2 ug/mL) for 24 h.Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). Anti-F4/80 rat monoclonal antibody was used to counterstain tubulin at 1/100 10 ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-CD204 antibody [EPR28270-88] - BSA and Azide free (AB314228)

This data was developed using ab314227, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) cells labelling CD204 with ab314227 at 1/50 (10.26 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing cytoplasmic staining in RAW 264.7 cell line.Negative control : Neuro-2a Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• Flow Cyt

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Flow Cytometry - Anti-CD204 antibody [EPR28270-88] - BSA and Azide free (AB314228)

This data was developed using ab314227, the same antibody clone in a different buffer formulation. Flow cytometric analysis of Neuro-2a (mouse neuroblastoma neuroblast, Left) / Raw264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage, Right) cells labelling CD204 with ab314227 at 1/500 dilution (0.1 ug)/Red compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody. Negative control : Neuro-2a.

• Flow Cyt

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Flow Cytometry - Anti-CD204 antibody [EPR28270-88] - BSA and Azide free (AB314228)

This data was developed using ab314227, the same antibody clone in a different buffer formulation. Flow cytometric analysis of Isotype (Left) / Mouse bone marrow treated with 2ug/ml LPS for 24h (Middle) / Untreated mouse bone marrow (Right) cells labelling CD204 with ab314227 at 1/5000 dilution (0.01 ug)/Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody. Cells are co-stained with F4/80 conjugated with Alexa Fluor®647. Gated on viable cells.

• WB

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Western blot - Anti-CD204 antibody [EPR28270-88] - BSA and Azide free (AB314228)

This data was developed using ab314227, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST Negative control : Neuro-2a, NIH/3T3. The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 35937296). In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution. Exposure time : Lane 1 : 5.5 seconds; Lane 2-4 : 37 seconds.

All lanes:

Western blot - Anti-CD204 antibody [EPR28270-88] (<a href='/en-us/products/primary-antibodies/cd204-antibody-epr28270-88-ab314227'>ab314227</a>) at 1/1000 dilution

Lane 1:

J774A.1 (mouse reticum cell sarcoma monocyte/macrophage ) whole cell lysate at 20 µg

Lane 2:

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

Lane 3:

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

Lane 4:

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='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 80 kDa

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