Anti-CD163 antibody [RM1114]

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-CD163 antibody [RM1114] (AB316218)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized THP-1 (human monocytic leukemia monocyte, Left) / SU-DHL-1 (human histiocytic lymphoblast-like cell, Right) cells labelling CD163 with ab316218 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 : THP-1.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD163 antibody [RM1114] (AB316218)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling CD163 with ab316218 at 1/500 (0.988 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on immune cells of human colon in (A) ab316218, non-specific staining on human colon in (B) ab87099.
The image on (A) ab316218 is applied with Anti-CD163 antibody at 1/500 dilution and (B) ab87099 is applied with Anti-CD163 antibody at 1/200 dilution.
The section was incubated with ab316218 or ab87099 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

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD163 antibody [RM1114] (AB316218)

Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling CD163 with ab316218 at 1/500 (0.988 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human spleen in (A) ab316218 and (B) ab87099.
The image on (A) ab316218 is applied with Anti-CD163 antibody at 1/500 dilution and (B) ab87099 is applied with Anti-CD163 antibody at 1/200 dilution.
The section was incubated with ab316218 or ab87099 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-CD163 antibody [RM1114] (AB316218)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized SU-DHL-1 (human histiocytic lymphoblast-like cell) cells labelling CD163 with ab316218 at 1/50 (9.88 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 SU-DHL-1 cell line.
Negative control : THP-1 (PMID : 10648003).
Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).
Left Panel is applied with Anti-CD163 antibody [RM1114] (ab316218) at 1/50 dilution;
Right panel is applied with Anti-CD163 antibody (ab87099) at 1/50 dilution.

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.

• IP

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Immunoprecipitation - Anti-CD163 antibody [RM1114] (AB316218)

CD163 was immunoprecipitated from 0.35 mg SU-DHL-1 (human histiocytic lymphoblast-like cell) whole cell lysate with ab316218 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab316218 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : SU-DHL-1 (human histiocytic lymphoblast-like cell) whole cell lysate
Lane 2 : ab316218 IP in SU-DHL-1 (human histiocytic lymphoblast-like cell) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab316218 in SU-DHL-1 whole cell lysate

All lanes:

Immunoprecipitation - Anti-CD163 antibody [RM1114] (ab316218) at 1/30 dilution

All lanes:

SU-DHL-1 (human histiocytic lymphoblast-like cell) whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>)

false

Exposure time: 15s

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD163 antibody [RM1114] (AB316218)

Immunohistochemical analysis of paraffin-embedded Rat spleen tissue labeling CD163 with ab316218 at 1/500 (0.988 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on rat spleen. The section was incubated with ab316218 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

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD163 antibody [RM1114] (AB316218)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling CD163 with ab316218 at 1/500 (0.988 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse spleen. The section was incubated with ab316218 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-CD163 antibody [RM1114] (AB316218)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling CD163 with ab316218 at 1/500 (0.988 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on kupffer cells of mouse liver. The section was incubated with ab316218 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

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Western blot - Anti-CD163 antibody [RM1114] (AB316218)

Left Panel is applied with Anti-CD163 antibody [RM1114] (ab316218) at 1/1000 dilution (0.494 ug/ml); Right panel is applied with Anti-CD163 antibody (ab87099) at 1/1000 dilution (0.2 ug/ml).

Exposure time : Lanes 1-2 : 180 seconds; Lane 3-4 : 26 seconds

All lanes:

Western blot - Anti-CD163 antibody [RM1114] (ab316218) at 1/1000 dilution

Lane 1:

Human spleen tissue lysate at 20 µg

Lane 2:

Human liver tissue lysate at 20 µg

Lane 3:

Human placenta tissue lysate at 20 µg

Lane 4:

SU-DHL-1 (human histiocytic lymphoblast-like cell) whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 150 kDa

false

• WB

Supplier Data

Western blot - Anti-CD163 antibody [RM1114] (AB316218)

Negative control : U-937 and THP-1(PMID : 10648003).

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

Exposure time : Lanes 1-2 : 180 seconds; Lane 3-6 : 26 seconds

All lanes:

Western blot - Anti-CD163 antibody [RM1114] (ab316218) at 1/1000 dilution

Lane 1:

Human spleen tissue lysate at 20 µg

Lane 2:

Human liver tissue lysate at 20 µg

Lane 3:

Human placenta tissue lysate at 20 µg

Lane 4:

SU-DHL-1 (human histiocytic lymphoblast-like cell) whole cell lysate at 20 µg

Lane 5:

U-937 (human histiocytic lymphoma monocyte) whole cell lysate at 20 µg

Lane 6:

THP-1 (human monocytic leukemia monocyte) whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 150 kDa,36 kDa

false