Anti-CD163 antibody [EPR19518] is a rabbit recombinant monoclonal antibody that is used to detect CD163 in Flow cytometry, IHC-Fr, IHC-P, Western blot, mIHC. Suitable for Human, Mouse, Rat samples.
- Cited in over 395 publications
- Using biophysical QC, antibody identity is confirmed at a molecular level for unrivaled batch-batch consistency
- Antibody clone EPR19518 is the most widely used clone for CD163 on the market
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Flow Cyt | WB | IHC-Fr | IHC-P | mIHC | |
---|---|---|---|---|---|
Human | Tested | Tested | Expected | Tested | Tested |
Mouse | Expected | Tested | Tested | Tested | Expected |
Rat | Expected | Tested | Expected | Tested | Expected |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/60 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/1000 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species Rat | Dilution info 1/1000 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species Human | Dilution info 1/1000 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/200 | Notes Antigen retrieval: Heated citrate solution (10mM citrate pH 6.0 + 0.05% Tween-20). |
Species | Dilution info | Notes |
---|---|---|
Species Human, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/500 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species Rat | Dilution info 1/500 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species Human | Dilution info 1/500 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/300 - 1/8000 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
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Acute phase-regulated receptor involved in clearance and endocytosis of hemoglobin/haptoglobin complexes by macrophages and may thereby protect tissues from free hemoglobin-mediated oxidative damage. May play a role in the uptake and recycling of iron, via endocytosis of hemoglobin/haptoglobin and subsequent breakdown of heme. Binds hemoglobin/haptoglobin complexes in a calcium-dependent and pH-dependent manner. Exhibits a higher affinity for complexes of hemoglobin and multimeric haptoglobin of HP*1F phenotype than for complexes of hemoglobin and dimeric haptoglobin of HP*1S phenotype. Induces a cascade of intracellular signals that involves tyrosine kinase-dependent calcium mobilization, inositol triphosphate production and secretion of IL6 and CSF1. Isoform 3 exhibits the higher capacity for ligand endocytosis and the more pronounced surface expression when expressed in cells. After shedding, the soluble form (sCD163) may play an anti-inflammatory role, and may be a valuable diagnostic parameter for monitoring macrophage activation in inflammatory conditions.
CD163, M130, Scavenger receptor cysteine-rich type 1 protein M130, Hemoglobin scavenger receptor
Anti-CD163 antibody [EPR19518] is a rabbit recombinant monoclonal antibody that is used to detect CD163 in Flow cytometry, IHC-Fr, IHC-P, Western blot, mIHC. Suitable for Human, Mouse, Rat samples.
- Cited in over 395 publications
- Using biophysical QC, antibody identity is confirmed at a molecular level for unrivaled batch-batch consistency
- Antibody clone EPR19518 is the most widely used clone for CD163 on the market
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Anti-CD163 antibody [EPR19518] (ab182422) was developed by Abcam using patented rabbit monoclonal antibody technology and is validated for use in Flow Cyt, IHC-Fr IHC-P, WB and mIHC.
Anti-CD163 antibody [EPR19518] (ab182422) was first used in a scientific publication in 2014 and has been cited over 399 times in peer reviewed journals. It's performance in IHC in human, mouse and rat samples is trusted by the scientific community.
Abcam's high quality manufacturing and validation processes ensure Anti-CD163 antibody [EPR19518] (ab182422) has high sensitivity and specificity alongside high lot-to-lot consistency and reproducibility.
Anti-CD163 antibody [EPR19518] (ab182422) has 17 independent reviews from customers.
Anti-CD163 antibody [EPR19518] (ab182422) specifically detects CD163 (UniProt ID: Q2VLH6; Molecular weight: 118kDa) and is sold in a convenient trial size to enable initial testing (20 µL) and larger sizes for subsequent scaling up experiments (100 µL and 1 mL).
Conjugation-ready, carrier free format available for antibody clone EPR19518 - Anti-CD163 antibody [EPR19518] - BSA and Azide free ab213612.
Antibody clone EPR19518 is also available pre-conjugated to a variety of labels for your convenience - Alexa Fluor® 555, Alexa Fluor® 594, Alexa Fluor® 488, Alexa Fluor® 647, Biotin, PE (Alexa Fluor® 555 Anti-CD163 antibody [EPR19518] ab281746, Alexa Fluor® 594 Anti-CD163 antibody [EPR19518] ab282114, Alexa Fluor® 488 Anti-CD163 antibody [EPR19518] ab313665, Alexa Fluor® 647 Anti-CD163 antibody [EPR19518] ab313666, Biotin Anti-CD163 antibody [EPR19518] ab314941, PE Anti-CD163 antibody [EPR19518] ab314947).
Recombinant antibody to CD163 is involved in conditions like rheumatoid arthritis and atherosclerosis, where it helps regulate inflammation. Top cited clone on the market
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
This product is a recombinant monoclonal antibody, which offers several advantages including:
For more information, read more on recombinant antibodies.
CD163 known also as hemoglobin scavenger receptor is a membrane glycoprotein with a molecular weight of approximately 130 kDa. It is predominantly expressed by monocytes and macrophages which are types of immune cells. Located on the cell surface CD163 functions as a receptor that aids in the clearance of hemoglobin-haptoglobin complexes playing a critical role in maintaining iron homeostasis and protecting tissues from oxidative damage. CD163 is not limited to human cells only; researchers often study its function using animal models such as chimeric rodents and chimeric rats.
CD163 participates in anti-inflammatory processes and is linked to immune modulation. It plays a pivotal role in the resolution of inflammation and promotes the removal of damaged cells and tissues. Through its involvement in the endocytic pathway CD163 interacts with various ligands facilitating interactions with other proteins. In the context of research CD163 functionality can be analyzed using techniques like CD163 IHC CD163 ELISA and CD163 stain in both human and mouse models.
CD163 is integral to the inflammation and immune response pathways. It participates in the heme-oxygenase pathway which is essential in the breakdown of heme into biliverdin free iron and carbon monoxide. This pathway involvement is tightly linked with heme oxygenase-1 (HO-1) a cytoprotective enzyme that helps to mitigate inflammatory responses. The interaction with haptoglobin and related proteins provides further insight into its regulatory roles within the immune system.
CD163 is associated with conditions like atherosclerosis and acute inflammation disorders. Its elevated levels indicate macrophage activation observed in chronic inflammations such as cardiovascular diseases. In atherosclerosis CD163 expression correlates with the uptake of oxidized LDL connecting it to progression of the disease. Additionally soluble CD163 levels serve as a biomarker in conditions like chronic liver disease where it is linked to macrophage activation and fibrosis. Within these disorders proteins like LDL and inflammatory cytokines are closely associated with CD163 function and regulation.
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Flow cytometry analysis of human PBMC cells (Human peripheral blood mononuclear cell) labeling with ab182422 at 1/60 dilution, 11.23 μg/ml (red). Goat anti rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) was used as the secondary antibody at 1/2000.
Formaldehyde-fixed, non-permeabilized human tonsil tissue stained for CD163 with ab182422 (30 mins at a 1/400 dilution) in immunohistochemical analysis. A Goat polyclonal HRP conjugate was used as the secondary.
Heat mediated antigen retrieval buffer/enzyme used: pH 9.0 EDTA.
Blocking step: 1% Protein Block ab64226 for 10 mins at RT.
Formaldehyde-fixed, non-permeabilized mouse liver tissue stained for CD163 with ab182422 (45 mins at a 1/400 dilution) in immunohistochemical analysis. A Rabbit polyclonal HRP conjugate was used as the secondary.
Heat mediated antigen retrieval buffer/enzyme used: pH 6.0 citrate.
Blocking step: 1% Protein Block ab64226 for 10 mins at RT.
Formaldehyde-fixed, non-permeabilized rat achilles tissue stained for CD163 with ab182422 (30 mins at a 1/200 dilution) in immunohistochemical analysis. A Rabbit polyclonal HRP conjugate was used as the secondary.
Heat mediated antigen retrieval buffer/enzyme used: pH 6.0 citrate 70°C for 2hrs.
Blocking step: 1% Protein Block ab64226 for 10 mins at RT.
10% NBF, non-permeabilized mouse spleen tissue stained for CD163 with ab182422 (12 hours, 4°C at a 1/200 dilution) in immunohistochemical analysis. A Donkey anti Rabbit IgG polyclonal AlexaFluor®647 conjugate was used as the secondary at a 1/200 dilution (red).
Heat mediated antigen retrieval buffer/enzyme used: Sodium citrate pH 6.0.
Blocking step: 5% serum for 1 hour at 21°C.
Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling CD163 with ab182422 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution. Cytoplasm staining on mouse spleen is observed. Counter stained with Hematoxylin. Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ab97051 at 1/500 dilution. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized frozen mouse spleen tissue labeling CD163 with ab182422 at 1/200 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution (green). The result showed some cytoplasmic staining on mouse spleen. The nuclear counterstain is DAPI (blue). Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 at 1/1000 dilution. Antigen retrieval: Heated citrate solution (10mM citrate pH 6.0 + 0.05% Tween-20).
Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1: 1 minute; Lane 2-5: 3 minutes.
U937 , THP-1 and J774A.1 cell lines were reported to be negative for CD163 expression.(PMID:16368951, 10648003 & 10577520).
The molecular weight observed is consistent with what has been described in the literature (PMID:9712057 & 16517975).
All lanes: Western blot - Anti-CD163 antibody [EPR19518] (ab182422) at 1/1000 dilution
Lane 1: Human fetal liver lysate at 20 µg
Lane 2: Human tonsil lysate at 20 µg
Lane 3: Human fetal spleen lysate at 20 µg
Lane 4: U937 (Human histiocytic lymphoma cell line) whole cell lysate at 20 µg
Lane 5: THP-1 (Human monocytic leukemia cell line) whole cell lysate at 20 µg
Lane 6: J774A.1 (Mouse macrophage reticulum cell sarcoma cell line) whole cell lysate at 20 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 125 kDa
Observed band size: 150 kDa
10% NBF, non-permeabilized rat muscle tissue stained for CD163 with ab182422 (12 hours, 4°C at a 1/200 dilution) in immunohistochemical analysis. A Donkey anti Rabbit IgG polyclonal AlexaFluor®647 conjugate was used as the secondary (red).
Heat mediated antigen retrieval buffer/enzyme used: Tris/EDTA pH 9.0.
Blocking step: 5% serum for 1 hour at 22°C.
Immunohistochemical analysis of paraffin-embedded human liver tissue labeling CD163 with ab182422 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution. Cytoplasm staining on Kupffer cells of human liver is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ab97051 at 1/500 dilution. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Formaldehyde-fixed, non-permeabilized human placenta tissue stained for CD163 with ab182422 (12 hours, 4°C at a 1/200 dilution) in immunohistochemical analysis. A Goat anti Rabbit polyclonal HRP conjugate was used as the secondary at a 1/200 dilution.
Heat mediated antigen retrieval buffer/enzyme used: pH 6.0 citrate buffer.
Blocking step: 3% serum for 30 mins at 20°C.
Different batches of ab182422 were tested on Rat liver lysate at 2.0 µg/ml. 15 µg of lysate was loaded in each lane. Bands observed at 150 kDa.
All lanes: Western blot - Anti-CD163 antibody [EPR19518] (ab182422)
Predicted band size: 125 kDa
Formaldehyde-fixed, non-permeabilized mouse liver tissue stained for CD163 with ab182422 (12 hours, 4°C at a 1/200 dilution) in immunohistochemical analysis. A Goat anti Rabbit HRP conjugate was used as the secondary at a 1/200 dilution.
Heat mediated antigen retrieval buffer/enzyme used: pH 6.0 citrate buffer.
Blocking step: 3% serum for 30 mins at 20°C.
10% Formalin-fixed, non-permeabilized human breast carcinoma tissue stained for CD163 with ab182422 (12 hours, 4°C at a 1/200 dilution) in immunohistochemical analysis. A Donkey anti Rabbit IgG polyclonal AlexaFluor®647 conjugate was used as the secondary at a 1/200 dilution (red).
Heat mediated antigen retrieval buffer/enzyme used: Tris/EDTA pH 9.0.
Blocking step: 5% serum for 1 hour at 22°C.
Formaldehyde-fixed, non-permeabilized human first trimester placenta tissue stained for CD163 with ab182422 (16 hours, 4°C at a 1/250 dilution) in immunohistochemical analysis. A Pig anti Rabbit polyclonal biotin conjugate was used as the secondary at a 1/250 dilution.
Heat mediated antigen retrieval buffer/enzyme used: Sodium citrate.
Blocking step: 5% BSA for 30 mins at 22°C.
10% NBF-fixed mouse spleen tissue stained for CD163 with ab182422 (18 hours at a 1/250 dilution) in immunohistochemical analysis. A Goat anti Rabbit IgG polyclonal AlexaFluor®647 conjugate was used as the secondary at a 1/600 dilution (red).
Heat mediated antigen retrieval buffer/enzyme used: Tris/EDTA pH 9.0.
Blocking step: 20% serum for 1 hour at RT.
Immunohistochemical analysis of paraffin-embedded human placenta tissue labeling CD163 with ab182422 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution. Cytoplasm staining on Hofbauer cells in human placenta is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ab97051 at 1/500 dilution. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemical analysis of paraffin-embedded mouse liver tissue labeling CD163 with ab182422 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution. Cytoplasm staining on Kupffer cells of mouse liver is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ab97051 at 1/500 dilution. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemical analysis of paraffin-embedded rat liver tissue labeling CD163 with ab182422 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution. Cytoplasm staining on Kupffer cells of rat liver is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ab97051 at 1/500 dilution. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized frozen mouse liver tissue labeling CD163 with ab182422 at 1/200 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution (green). The result showed some cytoplasmic staining on mouse liver. The nuclear counterstain is DAPI (blue).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 at 1/1000 dilution. Antigen retrieval: Heated citrate solution (10mM citrate pH 6.0 + 0.05% Tween-20).
Blocking/Dilution buffer: 5% NFDM/TBST.
The molecular weight observed is consistent with what has been described in the literature (PMID:9712057 & 16517975).
All lanes: Western blot - Anti-CD163 antibody [EPR19518] (ab182422) at 1/1000 dilution
Lane 1: Mouse liver lysate at 10 µg
Lane 2: Mouse heart lysate at 10 µg
Lane 3: Mouse spleen lysate at 10 µg
Lane 4: Mouse thymus lysate at 10 µg
Lane 5: Rat liver lysate at 10 µg
Lane 6: Rat heart lysate at 10 µg
Lane 7: Rat spleen lysate at 10 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 125 kDa
Observed band size: 150 kDa
Exposure time: 3min
Fluorescence multiplex immunohistochemical analysis of human liver (formalin-fixed paraffin-embedded section). Panel A shows merged staining of anti-MASP2 stained on cytoplasm of hepatocytes (Anti-MASP2 antibody [EPR23588-44] ab277520; gray; Opal™690) at 1:100 ( 5.22 μg/ml) [Panel B] , anti-CD163 stained on Kupffer cells (Anti-CD163 antibody [EPR19518] - BSA and Azide free ab213612; green; Opal™520) at 1:8000 ( 0.13 μg/ml) [Panel C], and anti-eNOS tained on endothelial cells (Anti-eNOS antibody [EPR23750-3] ab252439; red; Opal™570) at 1:200 ( 3.005 μg/ml) [Panel D] on human liver. DAPI was used as a nuclear counter stain. Followed by Opal Polymer HRP Ms + Rb secondary. 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. The section was incubated in three rounds of staining: in the order of Anti-MASP2 antibody [EPR23588-44] ab277520, Anti-CD163 antibody [EPR19518] - BSA and Azide free ab213612, and Anti-eNOS antibody [EPR23750-3] ab252439 for 30 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, epitope retrieval solution2) for 20 mins was used.
This data was developed using Anti-CD163 antibody [EPR19518] - BSA and Azide free ab213612, the same antibody clone in a different buffer formulation.
This data was developed using the same antibody clone in a different buffer formulation (Anti-CD163 antibody [EPR19518] - BSA and Azide free ab213612).
Immunohistochemical analysis of formalin fixed paraffin embedded Human tonsil labelling CD163 with Anti-CD163 antibody [EPR19518] - BSA and Azide free ab213612 at a concentration of 0.5 µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32mins.
Anti-CD163 antibody [EPR19518] - BSA and Azide free ab213612 anti CD163 antibody EPR19518 was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.
Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual)
Tissue Microarrays stained for "Anti-CD163 antibody [EPR19518]" using "ab182422"in immunohistochemical analysis. This table provides a detailed overview of positive (tick mark) and negative (cross mark) staining per sample type tested. The sections were pre-treated using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 minutes. The sections were incubated with ab182422 for 30 mins at room temperature followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
Immunohistochemical analysis of formalin fixed paraffin embedded human tonsil labelling CD163 with ab182422 at 2 µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with an OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was conducted for 32min with ULTRA cell conditioning solution (CC1 pH8.5). ab182422 anti CD163 antibody was incubated at 37°C for 16min. Sections were counterstained is with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.
Immunohistochemical analysis of formalin fixed paraffin embedded human tonsil labelling CD163 with ab182422 at 2 µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with an OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was conducted for 32min with ULTRA cell conditioning solution (CC1 pH8.5). ab182422 anti CD163 antibody was incubated at 37°C for 16min. Sections were counterstained is with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.
Tissue Microarrays stained for "Anti-CD163 antibody [EPR19518]" using "ab182422"in immunohistochemical analysis. This table provides a detailed overview of positive (tick mark) and negative (cross mark) staining per sample type tested. The sections were pre-treated using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 minutes. The sections were incubated with ab182422 for 30 mins at room temperature followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
Fluorescence multiplex immunohistochemical analysis of human liver (formalin-fixed paraffin-embedded section). Panel A shows merged staining of anti-eNOS stained on endothelial cells (Anti-eNOS antibody [EPR23750-3] ab252439; red; Opal™570) at 1:1000 ( 1.004 μg/ml) [Panel B], anti-CD163 stained on Kupffer cells (Anti-CD163 antibody [EPR19518] - BSA and Azide free ab213612; green; Opal™520) at 1:8000 ( 0.13 μg/ml) [Panel B], and anti-Glucose Transporter GLUT2 stained on membrane of hepatocytes (Anti-Glucose Transporter GLUT2 antibody [EPR22946-74] ab234440; gray; Opal™690) at 1:200 ( 3.005 μg/ml) [Panel D] on human liver. DAPI was used as a nuclear counter stain. Followed by Opal Polymer HRP Ms + Rb secondary. 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. The section was incubated in three rounds of staining: in the order of Anti-Glucose Transporter GLUT2 antibody [EPR22946-74] ab234440, Anti-CD163 antibody [EPR19518] - BSA and Azide free ab213612, and Anti-eNOS antibody [EPR23750-3] ab252439 for 30 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, epitope retrieval solution2) for 20 mins was used.
This data was developed using Anti-CD163 antibody [EPR19518] - BSA and Azide free ab213612, the same antibody clone in a different buffer formulation.
This data was developed using Anti-CD163 antibody [EPR19518] - BSA and Azide free ab213612, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver labelling GLUT2 with Anti-Glucose Transporter GLUT2 antibody [EPR22946-74] - BSA and Azide free ab260003 at 1/1000 (B), eNOS with Anti-eNOS antibody [RM1181] ab317582 at 1/500 dilution (C) and CD163 with Anti-CD163 antibody [EPR19518] - BSA and Azide free ab213612 at 1/8000 dilution (D). Opal Polymer HRP Ms + Rb was used as a secondary antibody, and Nuclear DNA was labeled with DAPI (shown in blue). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Panel A: merged staining of anti-GLUT2 (magenta; Opal™690), anti-eNOS (green; Opal™520) and anti-CD163 (gray; Opal™570) on human liver.
Panel B: anti-GLUT2 staining membrane of hepatocytes in human liver.
Panel C: anti-eNOS staining endothelium in human liver.
Panel D: anti-CD163 staining Kupffer cells in human liver.
The section was incubated in three rounds of staining: in the order of Anti-Glucose Transporter GLUT2 antibody [EPR22946-74] - BSA and Azide free ab260003, Anti-eNOS antibody [RM1181] ab317582 and Anti-CD163 antibody [EPR19518] - BSA and Azide free ab213612 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.
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