Anti-CD68 antibody [EPR20545]

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD68 antibody [EPR20545] (AB213363)

Multiplex immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver tissue.

Panel A : Merged staining of Collagen VI (ab182744; green) anti-CD68 (ab213363; red) and anti-Lamin B1 (ab229025; magenta).

Panel B : Anti-Collagen VI (green) stained on extracellular matrix.

Panel C : Anti-CD68 (red) stained on Kupffer cells.

Panel D : Anti-Lamin B1 (magenta) stained on nuclear envelope.

Key protocol steps : The section was incubated in three rounds of staining with ab182744 (1/1000 dilution) ab213363 (1/1000 dilution) and ab229025 (1/4000 dilution) for 30 mins at room temperature. Each round was followed by tyramide signal amplification with the appropriate fluorophore. Heat mediated antigen retrieval was used (Tris-EDTA buffer (pH 9.0 epitope retrieval solution 2) for 20 mins after every round of antibody/fluorophore staining.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

DAPI was used as a nuclear counter stain. A ready-to-use anti-Rabbit and Mouse Polymer HRP was used as a secondary.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD68 antibody [EPR20545] (AB213363)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Human tonsil tissue staining PD-L1 with ab205921 at a 1 : 500 (2.19 ug/ml) dilution; PD1 with ab237728 at 1 : 2000 (0.525 ug/ml) dilution and CD68 with ab213363 at 1 : 500 (1.26 ug/ml) dilution followed by a ready to use secondary antibody Opal Polymer HRP Ms + Rb.

Panel A : merged staining of anti-CD68 (magenta; Opal^™690), anti-PD-L1 (green; Opal^™520) and anti-PD1 (red; Opal^™570) on human tonsil.
Panel B : anti-PD-L1 stained on cells involved in T cell inhibition.
Panel C : anti-PD1 stained on antigen-stimulated T cells.
Panel D : anti-CD68 stained on macrophages.

The section was incubated in three rounds of staining : in the order of ab213363 and ab205921 for 30 mins, then ab237728 for 10 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.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD68 antibody [EPR20545] (AB213363)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human heart atherosclerosis tissue staining LOX 1 with ab325339 at a 1/100 (5.05 μg/ml) dilution, ab310335 anti-Neutrophil Elastase used at a 1/100 (5.20 μg/ml) dilution and ab213363 anti-CD68 used at a 1/500 (1.314 μg/ml) dilution.

Panel A : anti-LOX 1 (green; Opal™520), anti-Neutrophil Elastase (magenta; Opal™570), anti-CD68 (yellow; Opal™690) on human heart atherosclerosis.

Panel B : anti-LOX 1 staining neutrophils in human heart atherosclerosis.

Panel C : anti-Neutrophil Elastase staining neutrophils in human heart atherosclerosis.

Panel D : anti-CD68 staining on macrophages in human heart atherosclerosis.

Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab325339, ab310335 and ab213363 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.

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD68 antibody [EPR20545] (AB213363)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human liver tissue staining LOX 1 with ab325339 at a 1/100 (5.05 μg/ml) dilution, ab310335 anti-Neutrophil Elastase used at a 1/100 (5.20 μg/ml) dilution and ab213363 anti-CD68 used at a 1/500 (1.314 μg/ml) dilution.

Panel A : anti-LOX 1 (green; Opal™520), anti-Neutrophil Elastase (magenta; Opal™570), anti-CD68 (yellow; Opal™690) on human liver.

Panel B : anti-LOX 1 staining neutrophils in human liver.

Panel C : anti-Neutrophil Elastase staining neutrophils in human liver.

Panel D : anti-CD68 staining on macrophages in human liver.

Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab325339, ab310335 and ab213363 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.

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-CD68 antibody [EPR20545] (AB213363)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human liver tissue staining MARCO with ab323717 at a 1 : 200 (2.49 ug/ml) dilution, ab213363 anti-CD68 used at 1 : 500 (1.26 ug/ml) dilution and ab237721 anti-CD3 used at a 1 : 2000 (0.26 ug/ml) dilution.

Panel A : merged staining of anti-MARCO (green; Opal™520), anti-CD68 (magenta; Opal™570) and anti-CD3 (yellow; Opal™690) on human liver.
Panel B : anti-MARCO staining macrophages in human liver.
Panel C : ant-CD68 staining macrophages in human liver.
Panel D : ant-CD3 staining T lymphocytes in human liver.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab323717, ab213363 and ab237721 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.

Nuclear counter stain with DAPI.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD68 antibody [EPR20545] (AB213363)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human spleen tissue staining LOX 1 with ab325339 at a 1/100 (5.05 μg/ml) dilution, ab310335 anti-Neutrophil Elastase used at a 1/100 (5.20 μg/ml) dilution and ab213363 anti-CD68 used at a 1/500 (1.314 μg/ml) dilution.

Panel A : anti-LOX 1 (green; Opal™520), anti-Neutrophil Elastase (magenta; Opal™570), anti-CD68 (yellow; Opal™690) on human spleen.

Panel B : anti-LOX 1 staining neutrophils in human spleen.

Panel C : anti-Neutrophil Elastase staining neutrophils in human spleen.

Panel D : anti-CD68 staining on macrophages in human spleen.

Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab325339, ab310335 and ab213363 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.

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-CD68 antibody [EPR20545] (AB213363)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human lung tissue staining SFTPA1 with ab325321 at a 1 : 1000 (0.512 ug/ml) dilution, ab213363 anti-CD68 used at 1 : 500 (1.314 ug/ml) dilution and ab239904 anti-Aquaporin 5 used at a 1 : 500 (2.0 ug/ml) dilution.

Panel A : merged staining of anti-SFTPA1 (green; Opal™520), anti-CD68 (magenta; Opal™690) and anti-Aquaporin 5 (gray; Opal™570) on human lung.
Panel B : anti-SFTPA1 staining type 2 pneumonocytes in human lung.
Panel C : anti-CD68 staining macrophages in human lung.
Panel D : anti-Aquaporin 5 staining epithelium and type 1 pneumonocytes in human lung.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab325324, ab213363 and ab239904 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.

Nuclear counter stain with DAPI.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD68 antibody [EPR20545] (AB213363)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human spleen tissue labelling PDI with ab243644 at 1.02 μg/mL (B), PD-L 1 with ab213524 at 1/100 dilution (C) and CD68 with ab213363 at 1/300 dilution (D). Anti-Rabbit and Mouse Polymer HRP was used as a secondary antibody, and DAPI was used for a nuclear counter stain. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20mins. Heat mediated antigen retrieval (Leica ER2, PH9.0, 20 minutes) was used in between rounds of tyramide signal amplification to remove the antibodies from the previous round, to avoid any cross-reactivity.

Panel A : merged staining of anti- PD1 (green, Opal™520), anti- PD-L1 (red, Opal™570) and anti- CD68 (yellow, Opal™690).

Panel B : Anti- PD1 stained on antigen-stimulated T cells.

Panel C : anti- PD-L1 stained on cells involved in T cell inhibition

Panel D : anti-CD68 stained on macrophages.

The section was incubated in three rounds of staining : in the order of ab243644, ab213363 and ab213524 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.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD68 antibody [EPR20545] (AB213363)

Immunohistochemical analysis of paraffin-embedded human tonsil tissue, labeling CD68 with ab213363 at 1/8000 dilution, followed by Goat anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining on macrophages of human tonsil is observed (PMID : 19543531). Counter stained with hematoxylin. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat anti-Rabbit IgG H&L (HRP) Ready to use.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD68 antibody [EPR20545] (AB213363)

Panel A : merged staining of anti-CD68 (magenta; Opal™690), anti-CD3 epsilon (green; Opal™520) and anti-CD19 (red; Opal™570) on Formalin/PFA-fixed paraffin-embedded sections of human spleen. Secondary antibody was Opal Polymer HRP Ms + Rb, and couterstaining was with DAPI. Panel B : anti-CD3 epsilon stained on T cells with ab16669 at 1/500 dilution Panel C : anti-CD19 stained on B cells with ab237772 at 1/5000 dilution Panel D : anti-CD68 stained on macrophages with ab213363 1/500 dilution The section was incubated in three rounds of staining : in the order of ab213363 and ab16669 for 30 mins, then ab237772 for 10 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. 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.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD68 antibody [EPR20545] (AB213363)

Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling CD68 with ab213363 at 1/5000 dilution. No blocking step performed. Anti-Rabbit HRP polymer was used as the secondary detection system. Heat-mediated antigen retrieval was performed using EDTA based pH 9.0 buffer.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD68 antibody [EPR20545] (AB213363)

Immunohistochemical analysis of paraffin-embedded human liver tissue labeling CD68 with ab213363 at 1/8000 dilution, followed by Goat anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining on Kupffer cells of human liver is observed (PMID : 12118106). Counter stained with hematoxylin. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat anti-Rabbit IgG H&L (HRP) Ready to use.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-CD68 antibody [EPR20545] (AB213363)

Immunofluorescent analysis of 100% methanol-fixed U937 (human histiocytic lymphoma cell line) cells labeling CD68 with ab213363 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on U937 cells.

The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) (red) at 1/200 dilution.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-CD68 antibody [EPR20545] (AB213363)

Immunofluorescent analysis of 100% methanol-fixed THP-1 (human monocytic leukemia cell line) cells labeling CD68 with ab213363 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on THP-1 cells.

The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) (red) at 1/200 dilution.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD68 antibody [EPR20545] (AB213363)

Immunohistochemical analysis of paraffin-embedded human cervical carcinoma tissue labeling CD68 with ab213363 at 1/8000 dilution, followed by Goat anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining on macrophages of human cervical carcinoma is observed (PMID : 12118106). Counter stained with hematoxylin. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat anti-Rabbit IgG H&L (HRP) Ready to use.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD68 antibody [EPR20545] (AB213363)

Tissue Microarrays stained for "Anti-CD68 antibody [EPR20545]" using "ab213363"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 ab93684 (Tris/EDTA buffer, pH 9.0). The sections were incubated with ab213363 at +4°C overnight followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD68 antibody [EPR20545] (AB213363)

Fluorescence multiplex immunohistochemical analysis of the human duodenum (Formalin/PFA-fixed paraffin-embedded sections). Panel A : merged staining of anti-liver FABP (ab240401 gray; Opal™690), anti-CD3 epsilon (ab16669 green; Opal™520) and anti-CD68 (ab213363 red; Opal™570) on human duodenum. Panel B : anti-liver FABP stained on enterocytes. Panel C : anti-CD3 epsilon stained on T cells. Panel D : anti-CD68 stained on macrophages. Opal Polymer HRP Ms + Rb was used as a secondary antibody. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal™ 4-color kit. The section was incubated in three rounds of staining : in the order of ab240401 (1/8000 dilution), ab16669 (1/150 dilution) and ab213363 (1/500 dilution) 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. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Leica SP8 confocal microscope.

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-CD68 antibody [EPR20545] (AB213363)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Human thymus tissue labeling CD3 epsilon with ab16669 at 1/500 dilution, LY75/DEC-205 with ab208649 at 1/15000, and CD68 with ab213363 at 1/500 dilution. Panel A : merged staining of anti-CD68 (magenta; Opal™690), anti-CD3 epsilon (green; Opal™520) and anti-LY75/DEC-205 (red; Opal™570) on human thymus. Panel B : anti-CD3 epsilon stained on T cells. Panel C : anti-LY75/DEC-205 stained on thymic cortical epithelium and dendritic cells. Panel D : anti-CD68 stained on macrophages. Sections  were treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins before antibody incubation. The section was incubated in three rounds of staining : in the order of ab213363, ab16669, and ab208649 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. DAPI was used as a nuclear counterstain. 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.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD68 antibody [EPR20545] (AB213363)

Panel A : merged staining of anti-CD68 (gray; Opal™690), anti-CD3 epsilon (green; Opal™520) and anti-CD19 (red; Opal™570) on Formalin/PFA-fixed paraffin-embedded sections of human tonsil. Secondary antibody was Opal Polymer HRP Ms + Rb, and counterstaining was with DAPI. Panel B : anti-CD3 epsilon stained on T cells with ab16669 at 1/500 dilution Panel C : anti-CD19 stained on B cells with ab237772 at 1/5000 dilution Panel D : anti-CD68 stained on macrophages with ab213363 1/500 dilution The section was incubated in three rounds of staining : in the order of ab213363 and ab16669 for 30 mins, then ab237772 for 10 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. 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.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD68 antibody [EPR20545] (AB213363)

Fluorescence multiplex immunohistochemical analysis of the human colon (Formalin/PFA-fixed paraffin-embedded sections). Panel A : merged staining of anti-liver FABP (ab240401 gray; Opal™690), anti-CD3 epsilon (ab16669 green; Opal™520) and anti-CD68 (ab213363 red; Opal™570) on human colon. Panel B : anti-liver FABP stained on enterocytes. Panel C : anti-CD3 epsilon stained on T cells. Panel D : anti-CD68 stained on macrophages. Opal Polymer HRP Ms + Rb was used as a secondary antibody. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal™ 4-color kit. The section was incubated in three rounds of staining : in the order of ab240401 (1/8000 dilution), ab16669 (1/150 dilution) and ab213363 (1/500 dilution) 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. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Leica SP8 confocal microscope.

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-CD68 antibody [EPR20545] (AB213363)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human tonsil tissue staining PD1 with ab309363 at a 1/4000 dilution ( 0.125 μg/ml), ab228462 anti-PD-L1 used at 1/100 dilution (0.52 μg/ml) and ab213363 anti-CD68 used at a 1/500 dilution (1.26 μg/ml).

Panel A : merged staining of anti-CD68 (gray; Opal™690), anti-PD1 (green; Opal™520) and anti-PD-L1 (red; Opal™570) on human tonsil.
Panel B : anti-PD1 stained on antigen-stimulated T cells.
Panel C : anti-PD-L1 stained on cells involved in T cell inhibition.
Panel D : anti-CD68 stained on macrophages.

The section was incubated in three rounds of staining : in the order of ab213363 and ab309363 for 30 mins, then ab228462 for 10 mins at room temperature.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins. 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.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD68 antibody [EPR20545] (AB213363)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human placenta tissue staining CD68 with ab213363 at a 1/500 dilution, ab315802 anti-CD34 used at 1/2000 dilution and ab255296 anti-CD177 used at a 1/2000 dilution.

Panel A : merged staining of anti-CD68 (green; Opal™520), anti-CD34 (magenta; Opal™690) and anti-CD177 (gray; Opal™570) on human placenta.

Panel B : anti-CD68 staining macrophages in human placenta.

Panel C : ant-CD34 staining endothelium in human placenta.

Panel D : ant-CD177 staining neutrophils in human placenta.

Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab213363, ab315802 and ab255296 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.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD68 antibody [EPR20545] (AB213363)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human lung tissue staining SFTPA1 + SFTPA2 with ab320648 at a 1/1000 dilution, ab213363 anti-CD68 used at 1/500 dilution and ab307666 anti-Uteroglobin used at a 1/2000 dilution.

Panel A : merged staining of anti-SFTPA1+SFTPA2 (magenta; Opal^™690), anti-CD68 (green; Opal^™520) and anti-Uteroglobin (gray; Opal^™570) on human lung.
Panel B : anti-SFTPA1+SFTPA2 staining alveolar type II cells in human lung.
Panel C : ant-CD68 staining macrophages in human lung.
Panel D : ant-Uteroglobin staining club cells in human lung.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab320648, ab211363 and ab307666 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.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins..

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD68 antibody [EPR20545] (AB213363)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human liver cancer tissue staining CD5L with ab323268 at a 1/2000 dilution (0.246 μg/ml) and CD68 with ab213363 at 1/500 dilution (1.26 μg/ml), followed by a ready to use secondary antibody Opal Polymer HRP Ms + Rb.

Panel A : merged staining of anti-CD5L (green; Opal™520) and anti-CD68 (magenta; Opal™690) on human liver cancer.
Panel B : anti-CD5L staining Kupffer cells in human liver cancer.
Panel C : anti-CD68 staining Kupffer cells in human liver cancer.
Panel D : Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab323268 and ab213363 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.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD68 antibody [EPR20545] (AB213363)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human spleen tissue staining CD5L with ab323268 at a 1/2000 dilution (0.246 μg/ml) and CD68 with ab213363 at 1/500 dilution (1.26 μg/ml), followed by a ready to use secondary antibody Opal Polymer HRP Ms + Rb.

Panel A : merged staining of anti-CD5L (green; Opal™520) and anti-CD68 (magenta; Opal™690) on human spleen.
Panel B : anti-CD5L staining Kupffer cells in human spleen.
Panel C : anti-CD68 staining Kupffer cells in human spleen.
Panel D : Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab323268 and ab213363 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.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD68 antibody [EPR20545] (AB213363)

Immunohistochemical analysis of formalin fixed paraffin embedded Human liver labelling active CD68 with ab213363 at a concentration of 0.08 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.

ab213363 Anti CD68 antibody EPR20545 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)

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD68 antibody [EPR20545] (AB213363)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human liver tissue staining CD5L with ab323268 at a 1/2000 dilution (0.246 μg/ml) and CD68 with ab213363 at 1/500 dilution (1.26 μg/ml), followed by a ready to use secondary antibody Opal Polymer HRP Ms + Rb.

Panel A : merged staining of anti-CD5L (green; Opal™520) and anti-CD68 (magenta; Opal™690) on human liver.
Panel B : anti-CD5L staining Kupffer cells in human liver.
Panel C : anti-CD68 staining Kupffer cells in human liver.
Panel D : Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab323268 and ab213363 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.

• WB

Supplier Data

Western blot - Anti-CD68 antibody [EPR20545] (AB213363)

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure time : Lane 1/2/4/5 : 30 seconds; Lane 3 : 3 minutes.

The observed molecular weight is consistent with the literature (PMID : 18405323; PMID : 11739566; PMID : 16710801).

Lanes 1, 2, 3 and 5:

Western blot - Anti-CD68 antibody [EPR20545] (ab213363) at 1/1000 dilution

Lane 4:

Western blot - Anti-CD68 antibody [EPR20545] (ab213363) at 1/5000 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:

THP-1 (human monocytic leukemia cell line) whole cell lysate at 10 µg

Lane 5:

U937 (human histiocytic lymphoma cell line) whole cell lysate at 10 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

Predicted band size: 37 kDa

Observed band size: 110 kDa

true