Anti-Cytokeratin 19 antibody [EP1580Y] - BSA and Azide free

22 product images

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  Immunocytochemistry/ Immunofluorescence - Anti-Cytokeratin 19 antibody [EP1580Y] - BSA and Azide free (ab195872)

  Clone EP1580Y (ab195872) has been successfully conjugated by Abcam. This image was generated using Anti-Cytokeratin 19 antibody [EP1580Y] (Alexa Fluor® 488). Please refer to Alexa Fluor® 488 Anti-Cytokeratin 19 antibody [EP1580Y] ab192643 for protocol details.

  Alexa Fluor® 488 Anti-Cytokeratin 19 antibody [EP1580Y] ab192643 staining Cytokeratin 19 in wild-type MCF7 cells (top panel) and KRT19 knockout MCF7 cells (bottom panel). The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with Alexa Fluor® 488 Anti-Cytokeratin 19 antibody [EP1580Y] ab192643 at 1/500 dilution (shown in green) and Alexa Fluor® 647 Anti-Tubulin antibody [YOL1/34] - Microtubule Marker ab195884 (Rat monoclonal to Tubulin - Alexa Fluor® 647) at 1/100 dilution (shown in red) overnight at +4°C. Nuclear DNA was labelled in blue with DAPI.
  Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

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  Flow Cytometry (Intracellular) - Anti-Cytokeratin 19 antibody [EP1580Y] - BSA and Azide free (ab195872)

  Anti-Cytokeratin 19 antibody [EP1580Y] - Cytoskeleton Marker ab52625 staining Cytokeratin 19 in the human cell line MCF-7 (human breast carcinoma) by intracellular flow cytometry. Cells were fixed with 4% paraformaldehyde, permeabilized with 90% methanol and the sample was incubated with the primary antibody at a dilution of 1/80. A goat anti rabbit IgG (Alexa Fluorr^® 488) at a dilution of 1/2000 was used as the secondary antibody.
  

  Isoytype control: Rabbit monoclonal IgG (Black)

  Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Cytokeratin 19 antibody [EP1580Y] - Cytoskeleton Marker ab52625).

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  Immunocytochemistry/ Immunofluorescence - Anti-Cytokeratin 19 antibody [EP1580Y] - BSA and Azide free (ab195872)

  Clone EP1580Y (ab195872) has been successfully conjugated by Abcam. This image was generated using Anti-Cytokeratin 19 antibody [EP1580Y] (Alexa Fluor® 647). Please refer to Alexa Fluor® 647 Anti-Cytokeratin 19 antibody [EP1580Y] ab192980 for protocol details.

  Alexa Fluor® 647 Anti-Cytokeratin 19 antibody [EP1580Y] ab192980 staining Cytokeratin 19 in HeLa cells. The cells were fixed with 100% methanol (5 min) and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Triton X-100 for 1hr. The cells were then incubated with Alexa Fluor® 647 Anti-Cytokeratin 19 antibody [EP1580Y] ab192980 at a working dilution of 1 in 50 (shown in red) and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse monoclonal [DM1A] to alpha Tubulin) at 1µg/ml overnight at +4°C, followed by a further incubation at room temperature for 1hr with an AlexaFluor^® 488 Goat anti-mouse IgG (H&L - preadsorbed) secondary (Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.

  This product gave a positive signal in 4% formaldehyde (10 min) fixed HeLa cells under the same testing conditions

  Image was taken with a Confocal microscope (Leica micro-systems, TCS SP8).

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  Western blot - Anti-Cytokeratin 19 antibody [EP1580Y] - BSA and Azide free (ab195872)

  This data was developed using Anti-Cytokeratin 19 antibody [EP1580Y] - Cytoskeleton Marker ab52625, the same antibody clone in a different buffer formulation. Different batches of Anti-Cytokeratin 19 antibody [EP1580Y] - Cytoskeleton Marker ab52625 were tested on HepG2 (Human hepatocellular carcinoma epithelial cell) lysate at 0.2 µg/ml. 15 µg of lysate was loaded in each lane. Bands observed at 40 kDa.

  All lanes: Western blot - Anti-Cytokeratin 19 antibody [EP1580Y] - Cytoskeleton Marker (Anti-Cytokeratin 19 antibody [EP1580Y] - Cytoskeleton Marker ab52625)

  Predicted band size: 44 kDa

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  Immunocytochemistry/ Immunofluorescence - Anti-Cytokeratin 19 antibody [EP1580Y] - BSA and Azide free (ab195872)

  This data was developed using the same antibody clone in a different buffer formulation (Anti-Cytokeratin 19 antibody [EP1580Y] - Cytoskeleton Marker ab52625).
  

  Anti-Cytokeratin 19 antibody [EP1580Y] - Cytoskeleton Marker ab52625 staining Cytokeratin 19 in wild-type MCF7 cells (top panel) and KRT19 knockout MCF7 cells (bottom panel). The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with Anti-Cytokeratin 19 antibody [EP1580Y] - Cytoskeleton Marker ab52625 at 1/100 dilution and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor^® 594) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
  Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

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  Flow Cytometry (Intracellular) - Anti-Cytokeratin 19 antibody [EP1580Y] - BSA and Azide free (ab195872)

  Clone EP1580Y (ab195872) has been successfully conjugated by Abcam. This image was generated using Anti-Cytokeratin 19 antibody [EP1580Y] (PE). Please refer to PE Anti-Cytokeratin 19 antibody [EP1580Y] ab224981 for protocol details.
  

  Overlay histogram showing HeLa cells stained with PE Anti-Cytokeratin 19 antibody [EP1580Y] ab224981 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (PE Anti-Cytokeratin 19 antibody [EP1580Y] ab224981, 1/1000 dilution) for 30 min at 22°C.

  Isotype control antibody (black line) was Rabbit IgG (monoclonal) Phycoerythrin (PE Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab209478) used at the same concentration and conditions as the primary antibody.

  Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 50 mW Yellow/Green laser (561nm) and 586/15 bandpass filter.

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  Immunocytochemistry/ Immunofluorescence - Anti-Cytokeratin 19 antibody [EP1580Y] - BSA and Azide free (ab195872)

  Immunocytochemistry/Immunofluorescence analysis of HepG2 (human liver hepatocellular carcinoma cell line) cells labelling Cytokeratin 19 (green) with purified Anti-Cytokeratin 19 antibody [EP1580Y] - Cytoskeleton Marker ab52625 at 1/500. Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077, Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. Cells were counter-stained with Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, anti-Tubulin (mouse mAb) at 1/1000 followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 AlexaFluor^®594 goat anti-mouse secondary (1/1000). Nuclei were counterstained with DAPI (blue).

  For negative control 1, rabbit primary antibody and anti-mouse secondary antibody (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) were used. For negative control 2, Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (mouse primary antibody) was used followed by anti-rabbit secondary antibody (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077).  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Cytokeratin 19 antibody [EP1580Y] - Cytoskeleton Marker ab52625).

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  Immunocytochemistry/ Immunofluorescence - Anti-Cytokeratin 19 antibody [EP1580Y] - BSA and Azide free (ab195872)

  Immunofluorescent staining of MCF-7 cells (fixed in 4% PFA, permeabilized with 0.1% Triton X 100) using purified Anti-Cytokeratin 19 antibody [EP1580Y] - Cytoskeleton Marker ab52625 at a dilution of 1/200. An Alexa Fluor^® 488 goat anti-rabbit antibody was used as the secondary at a dilution of 1/200 and the cells were counter stained with DAPI.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Cytokeratin 19 antibody [EP1580Y] - Cytoskeleton Marker ab52625).

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 19 antibody [EP1580Y] - BSA and Azide free (ab195872)

  Immunohistochemical staining of paraffin-embedded human skin with purified Anti-Cytokeratin 19 antibody [EP1580Y] - Cytoskeleton Marker ab52625 at a dilution of 1/400. A pre-diluted HRP polymer for rabbit/mouse IgG was used as the secondary antibody and the sample was counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Cytokeratin 19 antibody [EP1580Y] - Cytoskeleton Marker ab52625).

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 19 antibody [EP1580Y] - BSA and Azide free (ab195872)

  Unpurified Anti-Cytokeratin 19 antibody [EP1580Y] - Cytoskeleton Marker ab52625 showing positive staining in Breast carcinoma tissue.
  
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Cytokeratin 19 antibody [EP1580Y] - Cytoskeleton Marker ab52625).

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 19 antibody [EP1580Y] - BSA and Azide free (ab195872)

  Unpurified Anti-Cytokeratin 19 antibody [EP1580Y] - Cytoskeleton Marker ab52625 showing positive staining in Endometrial carcinoma tissue.
  
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Cytokeratin 19 antibody [EP1580Y] - Cytoskeleton Marker ab52625).

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 19 antibody [EP1580Y] - BSA and Azide free (ab195872)

  Unpurified Anti-Cytokeratin 19 antibody [EP1580Y] - Cytoskeleton Marker ab52625 showing negative staining in Glioma tissue.
  
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Cytokeratin 19 antibody [EP1580Y] - Cytoskeleton Marker ab52625).

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 19 antibody [EP1580Y] - BSA and Azide free (ab195872)

  Unpurified Anti-Cytokeratin 19 antibody [EP1580Y] - Cytoskeleton Marker ab52625 showing positive staining in Gastric adenocarcinoma tissue.
  
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Cytokeratin 19 antibody [EP1580Y] - Cytoskeleton Marker ab52625).

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  Flow Cytometry (Intracellular) - Anti-Cytokeratin 19 antibody [EP1580Y] - BSA and Azide free (ab195872)

  Overlay histogram showing HeLa (human epithelial cell line from cervix adenocarcinoma) cells stained with unpurified Anti-Cytokeratin 19 antibody [EP1580Y] - Cytoskeleton Marker ab52625 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (Anti-Cytokeratin 19 antibody [EP1580Y] - Cytoskeleton Marker ab52625, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight^® 488 goat anti-rabbit IgG (H+L) (Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Cytokeratin 19 antibody [EP1580Y] - Cytoskeleton Marker ab52625).

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  OI-RD Scanning - Anti-Cytokeratin 19 antibody [EP1580Y] - BSA and Azide free (ab195872)

  We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody.
  Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.

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  Multiplex immunohistochemistry - Anti-Cytokeratin 19 antibody [EP1580Y] - BSA and Azide free (ab195872)

  Fluorescence multiplex immunohistochemical analysis of human liver tissue (formalin-fixed paraffin-embedded section). Panel A shows merged staining of ab195872 anti-Cytokeratin 19 stained on branch of bile ducts (magenta; Opal™690) at 1:8000 ( 0.127 μg/ml) [Panel B], Anti-Factor VIII antibody [EPR24039-262] ab275376 anti-Factor VIII stained on endothelial cells (red; Opal™570) at 1:1000 ( 0.457 μg/ml) [Panel C], and Anti-alpha smooth muscle Actin antibody [SP171] ab150301 anti-alpha smooth muscle Actin stained on smooth muscles (green; Opal™520) at 1:200 ( 0.14 μg/ml) [Panel C] 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 ab195872 for 30 mins, Anti-Factor VIII antibody [EPR24039-262] ab275376 for 30 mins and Anti-alpha smooth muscle Actin antibody [SP171] ab150301 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 was used. 

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  Multiplex immunohistochemistry - Anti-Cytokeratin 19 antibody [EP1580Y] - BSA and Azide free (ab195872)

  Cytokeratin 19 Multiplex immunohistochemistry staining of Mouse liver using rabbit Anti-Cytokeratin 19 antibody

  This data was developed using Anti-Cytokeratin 19 antibody [EP1580Y] - Cytoskeleton Marker ab52625, the same antibody clone in a different buffer formulation.
  

  Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse liver tissue staining COL4A2 with Anti-COL4A2 antibody [EPR25363-49] ab316099 at a 1/500 dilution, Anti-ALDH1L1 antibody [EPR25443-103] - Astrocyte Marker ab307696 anti-ALDH1L1 used at 1/5000 dilution and Anti-Cytokeratin 19 antibody [EP1580Y] - Cytoskeleton Marker ab52625 anti-Cytokeratin 19 used at a 1/6000 dilution.

  Panel A: merged staining of anti-COL4A2 (green; Opal^™520), anti-ALDH1L1 (magenta; Opal^™690) and anti-Cytokeratin 19 (grey; Opal^™570) on mouse liver.
  
  Panel B: anti-COL4A2 staining endothelium in mouse liver.
  
  Panel C: anti-ALDH1L1 staining hepatocytes in mouse liver.
  
  Panel D: anti-Cytokeratin 19 staining branch of bile ducts in mouse liver.

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

  The section was incubated in three rounds of staining: in the order of Anti-COL4A2 antibody [EPR25363-49] ab316099, Anti-ALDH1L1 antibody [EPR25443-103] - Astrocyte Marker ab307696 and Anti-Cytokeratin 19 antibody [EP1580Y] - Cytoskeleton Marker ab52625 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.

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  Multiplex immunohistochemistry - Anti-Cytokeratin 19 antibody [EP1580Y] - BSA and Azide free (ab195872)

  Cytokeratin 19 Multiplex immunohistochemistry staining of Human liver tissue using rabbit Anti-Cytokeratin 19 antibody

  This data was developed using Anti-Cytokeratin 19 antibody [EP1580Y] - Cytoskeleton Marker ab52625, the same antibody clone in a different buffer formulation.
  

  Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human liver tissue staining Carboxypeptidase A with Anti-VAP1 antibody [EPR28748-59] ab317620 at a 1/1000 dilution, Anti-Inhibin beta E chain antibody [EPR29191-125] ab322202 anti-Inhibin beta E chain used at 1/2000 dilution and Anti-Cytokeratin 19 antibody [EP1580Y] - Cytoskeleton Marker ab52625 anti-Cytokeratin 19 used at a 1/6000 dilution. Opal Polymer HRP Ms + Rb was used as a secondary antibody.
  

  Panel A: merged staining of anti-VAP1 (green; Opal^™520), anti-Inhibin beta E chain (magenta; Opal^™690) and anti-Cytokeratin 19 (gray; Opal^™570) on human liver.
  Panel B: anti-VAP1 staining endothelium in human liver.
  
  Panel C: anti-Inhibin beta E chain staining hepatocytes in human liver.
  
  Panel D: anti-Cytokeratin 19 staining branch of bile ducts 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 Anti-VAP1 antibody [EPR28748-59] ab317620, Anti-Inhibin beta E chain antibody [EPR29191-125] ab322202 and Anti-Cytokeratin 19 antibody [EP1580Y] - Cytoskeleton Marker ab52625 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.

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  Flow Cytometry (Intracellular) - Anti-Cytokeratin 19 antibody [EP1580Y] - BSA and Azide free (ab195872)

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Cytokeratin 19 antibody [EP1580Y] - Cytoskeleton Marker ab52625).

  Flow cytometry overlay histogram showing left MCF7 positive cells and right negative SH-SY5Y stained with Anti-Cytokeratin 19 antibody [EP1580Y] - Cytoskeleton Marker ab52625 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 mins. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (Anti-Cytokeratin 19 antibody [EP1580Y] - Cytoskeleton Marker ab52625) (1x 10⁶ in 100μl at 0.04μg/ml (1/50,000 dilution)) for 30 mins at 22°C.

  The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed was incubated at 1/4000 dilutionfor 30 mins at 22°C

  Isotype control antibody (black line) was Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

  Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.

  This antibody gave a positive signal in MCF7 Fixed with 80% methanol (5 min) / permeabilised with 0.1% PBS-Triton X-100 for 15 mins under the same conditions.

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  Multiplex immunohistochemistry - Anti-Cytokeratin 19 antibody [EP1580Y] - BSA and Azide free (ab195872)

  Fluorescence multiplex immunohistochemical analysis of the human pancreas (Formalin/PFA-fixed paraffin-embedded sections).
  
  Panel A: merged staining of anti-Carboxypeptidase A (Anti-Carboxypeptidase A antibody [EPR24384-69] ab278044, magenta; Opal™690), anti-Cytokeratin 19 (ab195872, green; Opal™520) and anti-Insulin (Anti-Insulin antibody [EPR17359] ab181547, red; Opal™570) on human pancreas. Panel B: anti-Carboxypeptidase A stained on acinar cells. Panel C: anti-Cytokeratin 19 stained on centroacinar cells and ducts. Panel D: anti-Insulin stained on beta cells. Opal Polymer HRP Ms + Rb was used as a secondary antibody.
  
  The section was incubated in three rounds of staining: in the order of Anti-Carboxypeptidase A antibody [EPR24384-69] ab278044 at 1/4000 dilution (0.135 μg/ml), ab195872 at 1/8000 dilution (0.127 μg/ml), and Anti-Insulin antibody [EPR17359] ab181547 at 1/20000 dilution (0.053 μg/ml) 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. DAPI (blue) was used as a nuclear counter stain.

• 

  Immunocytochemistry/ Immunofluorescence - Anti-Cytokeratin 19 antibody [EP1580Y] - BSA and Azide free (ab195872)

  Anti-Cytokeratin 19 antibody [EP1580Y] - Cytoskeleton Marker ab52625 staining KRT19 in MCF7 cells, with negative expression in SHSY5Y cells. The cells were fixed with 100% methanol (5 min), permeabilised with 0.1% Triton x-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with Anti-Cytokeratin 19 antibody [EP1580Y] - Cytoskeleton Marker ab52625 at 2 μg/ml and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, Mouse monoclonal [DM1A] to alpha Tubulin at 0.5 μg/ml. Cells were then incubated with Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 647) preadsorbed ab150119, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 647), pre-adsorbed at 1/1000 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue). Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown. This product also work with 4% formaldehyde (10 min) fixation under the same testing conditions.
  This data was developed using the same antibody clone in a different buffer formulation (Anti-Cytokeratin 19 antibody [EP1580Y] - Cytoskeleton Marker ab52625).

• 

  Multiplex immunohistochemistry - Anti-Cytokeratin 19 antibody [EP1580Y] - BSA and Azide free (ab195872)

  Cytokeratin 19 Multiplex immunohistochemistry staining of Rat liver using rabbit Anti-Cytokeratin 19 antibody

  This data was developed using Anti-Cytokeratin 19 antibody [EP1580Y] - Cytoskeleton Marker ab52625, the same antibody clone in a different buffer formulation.
  

  Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded rat liver tissue staining COL4A2 with Anti-COL4A2 antibody [EPR25363-49] ab316099 at a 1/500 dilution, Anti-ALDH1L1 antibody [EPR25443-103] - Astrocyte Marker ab307696 anti-ALDH1L1 used at 1/5000 dilution and Anti-Cytokeratin 19 antibody [EP1580Y] - Cytoskeleton Marker ab52625 anti-Cytokeratin 19 used at a 1/6000 dilution.

  Panel A: merged staining of anti-COL4A2 (green; Opal^™520), anti-ALDH1L1 (magenta; Opal^™690) and anti-Cytokeratin 19 (grey; Opal^™570) on rat liver.
  
  Panel B: anti-COL4A2 staining endothelium in rat liver.
  
  Panel C: anti-ALDH1L1 staining hepatocytes in rat liver.
  
  Panel D: anti-Cytokeratin 19 staining branch of bile ducts in rat liver.

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

  The section was incubated in three rounds of staining: in the order of Anti-COL4A2 antibody [EPR25363-49] ab316099, Anti-ALDH1L1 antibody [EPR25443-103] - Astrocyte Marker ab307696 and Anti-Cytokeratin 19 antibody [EP1580Y] - Cytoskeleton Marker ab52625 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.