Anti-CD68 antibody [EPR20545]

17 product images

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  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 (Anti-Collagen VI antibody [EPR17072] ab182744; green), anti-CD68 (ab213363; red) and anti-Lamin B1 (Anti-Lamin B1 antibody [EPR22165-121] 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 Anti-Collagen VI antibody [EPR17072] ab182744 (1/1000 dilution), ab213363 (1/1000 dilution) and Anti-Lamin B1 antibody [EPR22165-121] 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.

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  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 Anti-PD1 antibody [EPR23119-111] ab243644 at 1.02 μg/mL (B), PD-L 1 with Anti-PD-L1 antibody [EPR19759] 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 Anti-PD1 antibody [EPR23119-111] ab243644, ab213363 and Anti-PD-L1 antibody [EPR19759] 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.

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  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) (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) (Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker 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) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution.

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  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.

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  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) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Developed using the ECL technique.

  Predicted band size: 37 kDa

  Observed band size: 110 kDa

• 

  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) (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) (Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker 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) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution.

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  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.

• 

  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.

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  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.

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  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 Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) 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).

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  Multiplex immunohistochemistry - Anti-CD68 antibody [EPR20545] (ab213363)

  CD68 immunohistochemistry staining of human colon using rabbit anti-CD68 antibody
  
  

  Fluorescence multiplex immunohistochemical analysis of the human colon (Formalin/PFA-fixed paraffin-embedded sections). Panel A: merged staining of anti-liver FABP (Anti-liver FABP antibody [EPR20464] - BSA and Azide free ab240401 gray; Opal™690), anti-CD3 epsilon (Anti-CD3 epsilon antibody [SP7] 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 Anti-liver FABP antibody [EPR20464] - BSA and Azide free ab240401 (1/8000 dilution), Anti-CD3 epsilon antibody [SP7] 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.

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  Multiplex immunohistochemistry - Anti-CD68 antibody [EPR20545] (ab213363)

  CD68 immunohistochemistry staining of human duodenum using rabbit anti-CD68 antibody
  
  

  Fluorescence multiplex immunohistochemical analysis of the human duodenum (Formalin/PFA-fixed paraffin-embedded sections). Panel A: merged staining of anti-liver FABP (Anti-liver FABP antibody [EPR20464] - BSA and Azide free ab240401 gray; Opal™690), anti-CD3 epsilon (Anti-CD3 epsilon antibody [SP7] 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 Anti-liver FABP antibody [EPR20464] - BSA and Azide free ab240401 (1/8000 dilution), Anti-CD3 epsilon antibody [SP7] 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.

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  Multiplex immunohistochemistry - Anti-CD68 antibody [EPR20545] (ab213363)

  Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Human tonsil tissue staining PD-L1 with Anti-PD-L1 antibody [28-8] ab205921 at a 1:500 (2.19 ug/ml) dilution; PD1 with Anti-PD1 antibody [CAL20] 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 Anti-PD-L1 antibody [28-8] ab205921 for 30 mins, then Anti-PD1 antibody [CAL20] 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

• 

  Multiplex immunohistochemistry - Anti-CD68 antibody [EPR20545] (ab213363)

  Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human tonsil tissue staining PD1 with Anti-PD1 antibody [RM1033] ab309363 at a 1/4000 dilution ( 0.125 μg/ml), Anti-PD-L1 antibody [SP142] - C-terminal 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 Anti-PD1 antibody [RM1033] ab309363 for 30 mins, then Anti-PD-L1 antibody [SP142] - C-terminal 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.

• 

  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 Anti-CD3 epsilon antibody [SP7] ab16669 at 1/500 dilution
  Panel C: anti-CD19 stained on B cells with Anti-CD19 antibody [SP291] - BSA and Azide free 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 Anti-CD3 epsilon antibody [SP7] ab16669 for 30 mins, then Anti-CD19 antibody [SP291] - BSA and Azide free 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.

• 

  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 Anti-CD3 epsilon antibody [SP7] ab16669 at 1/500 dilution
  Panel C: anti-CD19 stained on B cells with Anti-CD19 antibody [SP291] - BSA and Azide free 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 Anti-CD3 epsilon antibody [SP7] ab16669 for 30 mins, then Anti-CD19 antibody [SP291] - BSA and Azide free 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.

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  Multiplex immunohistochemistry - Anti-CD68 antibody [EPR20545] (ab213363)

  CD68 immunohistochemistry staining of human thymus using rabbit anti-CD68 antibody
  
  Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Human thymus tissue labeling CD3 epsilon with Anti-CD3 epsilon antibody [SP7] ab16669 at 1/500 dilution, LY75/DEC-205 with Anti-LY75/DEC-205 antibody [EPR5233] - BSA and Azide free 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, Anti-CD3 epsilon antibody [SP7] ab16669, and Anti-LY75/DEC-205 antibody [EPR5233] - BSA and Azide free 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.