CD163 Recombinant Rabbit Monoclonal Antibody [JA51-30]

Fluorescence multiplex immunohistochemical analysis of the human cervical cancer (Formalin/PFA-fixed paraffin-embedded sections). Panel A: the merged image of anti-CD14 (ET1610-85, red), anti-S100A9 (ET1702-73, green), anti-CD68 (HA601115, cyan), anti-panCK (HA601138, magenta) and anti-CD163 (ET1704-43, yellow) on human cervical cancer. Panel B: anti- CD14 stained on monocyte and MDSCs. Panel C: anti-S100A9 stained on MDSCs. Panel D: anti-CD68 stained on macrophage M1 and macrophage M2. Panel E: anti-panCK stained on tumor cells. Panel F: anti-CD163 stained on macrophage M2. HRP Conjugated UltraPolymer Goat Polyclonal Antibody HA1119/HA1120 was used as a secondary antibody. The immunostaining was performed with the Sequential Immuno-staining Kit (IRISKit™MH010101, www.luminiris.cn). The section was incubated in five rounds of staining: in the order of ET1610-85 (1/1,000 dilution), ET1702-73 (1/1,000 dilution), HA601115 (1/2,000 dilution), HA601138 (1/3,000 dilution), and ET1704-43 (1/2,000 dilution) for 20 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 30 mins at 95℃. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Olympus VS200 Slide Scanner.

Fluorescence multiplex immunohistochemical analysis of the human pancreatic carcinoma (Formalin/PFA-fixed paraffin-embedded sections). Panel A: the merged image of anti-CD68 (EM1901-95, green), anti-CD163 (ET1704-43, red) and anti-PanCK (HA601094, violet) on human pancreatic carcinoma. Panel B: anti- CD68 stained on M1 macrophages. Panel C: anti-CD163 stained on M2 macrophages cells. Panel D: anti-panCK stained on cancer cells. HRP Conjugated UltraPolymer Goat Polyclonal Antibody HA1119/HA1120 was used as a secondary antibody. The immunostaining was performed with the Sequential Immuno-staining Kit (IRISKitCmTSA Kit 900803). The section was incubated in three rounds of staining: in the order of EM1901-95 (1/3,000 dilution), ET1704-43 (1/3,000 dilution), and HA601094 (1/3,000 dilution) for 20 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 30 mins at 95℃. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Nikon ECLIPSE Ni-E microscope.

Fluorescence multiplex immunohistochemical analysis of human cervical carcinoma (Formalin/PFA-fixed paraffin-embedded sections). Panel A: the merged image of anti-S100A9 (ET1702-73, White), anti-CD117 (HA721154, Red) and anti-CD163(ET1704-43, Yellow) on human cervical carcinoma. HRP Conjugated UltraPolymer Goat Polyclonal Antibody HA1119/HA1120 was used as a secondary antibody. The immunostaining was performed with the Sequential Immuno-staining Kit (IRISKit™MH010101, www.luminiris.cn). The section was incubated in three rounds of staining: in the order of ET1702-73 (1/1,000 dilution), HA721154 (1/1,000 dilution) and ET1704-43 (1/2,000 dilution) for 20 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 30 mins at 95℃. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Zeiss Observer 7 Inverted Fluorescence Microscope.

Western blot analysis of CD163 on different lysates with Rabbit anti-CD163 antibody (ET1704-43) at 1/1,000 dilution.

Lane 1: Human lung tissue lysate
Lane 2: Human liver tissue lysate

Lysates/proteins at 40 µg/Lane.

Predicted band size: 125 kDa
Observed band size: 150-170 kDa

Exposure time: 1 minute; ECL: K1801;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1704-43) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-CD163 antibody (ET1704-43) at 1/10,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1704-43) at 1/10,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-CD163 antibody (ET1704-43) at 1/10,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1704-43) at 1/10,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Immunohistochemical analysis of paraffin-embedded human placenta tissue with Rabbit anti-CD163 antibody (ET1704-43) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1704-43) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-CD163 antibody (ET1704-43) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1704-43) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Application: IF-Tissue

Species: Human

Site: Spleen

Sample: Paraffin-embedded section

Antibody concentration: 1/200