Anti-Cytokeratin 19 antibody [EP1580Y] - Cytoskeleton Marker

20 product images

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  Flow Cytometry (Intracellular) - Anti-Cytokeratin 19 antibody [EP1580Y] - Cytoskeleton Marker (ab52625)

  ab52625 staining Cytokeratin 19in 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).

  Alexa Fluorr^® 488 (Alexa Fluor® 488 Anti-Cytokeratin 19 antibody [EP1580Y] ab192643) and Alexa Fluorr^® 647 (Alexa Fluor® 647 Anti-Cytokeratin 19 antibody [EP1580Y] ab192980) conjugated versions are available for this clone.

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

  Blocking buffer: 5% NFDM/TBST

  Dilution buffer: 5% NFDM/TBST

  All lanes: Western blot - Anti-Cytokeratin 19 antibody [EP1580Y] - Cytoskeleton Marker (ab52625) at 1/45000 dilution

  Lane 1: HepG2 (liver hepatocellular carcinoma cell line) cell lysate at 20 µg

  Lane 2: NIH/3T3 (mouse embyro fibroblast cell line) cell lysate at 20 µg

  Secondary

  All lanes: HRP goat anti-rabbit (H+L) at 1/1000 dilution

  Predicted band size: 44 kDa

  Observed band size: 40 kDa

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

  Different batches of 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 (ab52625)

  Predicted band size: 44 kDa

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  Immunocytochemistry/ Immunofluorescence - Anti-Cytokeratin 19 antibody [EP1580Y] - Cytoskeleton Marker (ab52625)

  Immunocytochemistry/Immunofluorescence analysis of HepG2 (human liver hepatocellular carcinoma cell line) cells labelling Cytokeratin 19 (green) with purified 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 Alexa Fluor^® 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).

  Alexa Fluor^® 488 (Alexa Fluor® 488 Anti-Cytokeratin 19 antibody [EP1580Y] ab192643) and Alexa Fluor^® 647 (Alexa Fluor® 647 Anti-Cytokeratin 19 antibody [EP1580Y] ab192980) conjugated versions are available for this clone.

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  Immunocytochemistry/ Immunofluorescence - Anti-Cytokeratin 19 antibody [EP1580Y] - Cytoskeleton Marker (ab52625)

  Immunofluorescent staining of MCF-7 cells (fixed in 4% PFA, permeabilized with 0.1% Triton X 100) using purified 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.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 19 antibody [EP1580Y] - Cytoskeleton Marker (ab52625)

  Immunohistochemical staining of paraffin-embedded human skin with purified 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.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 19 antibody [EP1580Y] - Cytoskeleton Marker (ab52625)

  Unpurified ab52625 showing positive staining in Breast carcinoma tissue.
  
  
  

  Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 19 antibody [EP1580Y] - Cytoskeleton Marker (ab52625)

  Unpurified ab52625 showing positive staining in Endometrial carcinoma tissue.
  
  
  

  Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 19 antibody [EP1580Y] - Cytoskeleton Marker (ab52625)

  Unpurified ab52625 showing negative staining in Glioma tissue.
  
  
  

  Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 19 antibody [EP1580Y] - Cytoskeleton Marker (ab52625)

  Unpurified ab52625 showing positive staining in Gastric adenocarcinoma tissue.
  
  
  

  Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

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  Flow Cytometry (Intracellular) - Anti-Cytokeratin 19 antibody [EP1580Y] - Cytoskeleton Marker (ab52625)

  Overlay histogram showing HeLa (human epithelial cell line from cervix adenocarcinoma) cells stained with unpurified 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 (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.
  

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  OI-RD Scanning - Anti-Cytokeratin 19 antibody [EP1580Y] - Cytoskeleton Marker (ab52625)

  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] - Cytoskeleton Marker (ab52625)

  Fluorescence multiplex immunohistochemical analysis of human liver tissue (formalin-fixed paraffin-embedded section). Panel A shows merged staining of Anti-Cytokeratin 19 antibody [EP1580Y] - BSA and Azide free 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 Anti-Cytokeratin 19 antibody [EP1580Y] - BSA and Azide free 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. 
  
  This data was developed using Anti-Cytokeratin 19 antibody [EP1580Y] - BSA and Azide free ab195872, the same antibody clone in a different buffer formulation. 

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 19 antibody [EP1580Y] - Cytoskeleton Marker (ab52625)

  Immunohistochemical analysis of formalin fixed paraffin embedded human pancreas labelling Cytokeratin 19 with ab52625 at a concentration of 0.1µg/ml. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with a Bond™ Polymer Refine Detection kit. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution 2) for 20mins.
  ab52625 anti-Cytokeratin 19 antibody [EP1580Y] was incubated for 15mins at room temperature. Sections were counterstained with Hematoxylin. 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).

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 19 antibody [EP1580Y] - Cytoskeleton Marker (ab52625)

  Immunohistochemical analysis of formalin fixed paraffin embedded human pancreas labelling Cytokeratin 19 with ab52625 at a concentration of 0.1µg/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.
  ab52625 anti-Cytokeratin 19 antibody [EP1580Y] 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).

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  This image is courtesy of TissueGnostics Asia Pacific Limited

  Multiplex immunohistochemistry - Anti-Cytokeratin 19 antibody [EP1580Y] - Cytoskeleton Marker (ab52625)

  10-color fluorescence multiplex immunohistochemical analysis of human lung cancer tissue (formalin-fixed paraffin-embedded section).
  

  Merged staining of anti-FOXP3 (Anti-FOXP3 antibody [EPR22102-37] ab215206; Cyan; TG540N), anti-PD1 (Anti-PD1 antibody [NAT105] ab52587; Violet; TG700N), anti-CD163 (Anti-CD163 antibody [EPR19518] ab182422; Red; TG650N), anti-HLA-DR (Anti-HLA-DR antibody [EPR3692] ab92511; Yellow; TG570N), anti-CD4 (Anti-CD4 antibody [EPR6855] ab133616; Orange; TG620N), anti-CD8 alpha (Anti-CD8 alpha antibody [SP16] ab101500; Purple; TG540S), anti-CD20 (Anti-CD20 antibody [L26] ab9475; Grey; TG660S), anti-CD68 (Anti-CD68 antibody [SP251] ab192847; Green; TG520N), anti-Cytokeratin 19 (ab52625; Light blue; TG440N). TG470SN (dark blue) was used as a nuclear counter stain.

  The section was incubated in nine rounds of staining; in the order of Anti-FOXP3 antibody [EPR22102-37] ab215206 (1/100 dilution), Anti-PD1 antibody [NAT105] ab52587 (1/200 dilution), Anti-CD163 antibody [EPR19518] ab182422 (1/300 dilution), Anti-HLA-DR antibody [EPR3692] ab92511 (1/200 dilution), Anti-CD4 antibody [EPR6855] ab133616 (1/600 dilution), Anti-CD8 alpha antibody [SP16] ab101500 (1/300 dilution), Anti-CD20 antibody [L26] ab9475 (1/100 dilution), Anti-CD68 antibody [SP251] ab192847 (1/300 dilution), ab52625 (1/400 dilution); each using a separate fluorescent tyramide signal amplification system.

  Sodium citrate antigen retrieval (pH6.0) was used in between rounds of tyramide signal amplification to remove the antibody from the previous round, to avoid any cross-reactivity.

  Image acquisition was performed with TissueFAXS Spectra (TissueGnostics).

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  This image is courtesy of TissueGnostics Asia Pacific Limited

  Multiplex immunohistochemistry - Anti-Cytokeratin 19 antibody [EP1580Y] - Cytoskeleton Marker (ab52625)

  10-color fluorescence multiplex immunohistochemical analysis of human lung cancer tissue (formalin-fixed paraffin-embedded section).

  Merged and separate images on the staining of anti-FOXP3 (Anti-FOXP3 antibody [EPR22102-37] ab215206), anti-PD1 (Anti-PD1 antibody [NAT105] ab52587), anti-CD163 (Anti-CD163 antibody [EPR19518] ab182422), anti-HLA-DR (Anti-HLA-DR antibody [EPR3692] ab92511), anti-CD4 (Anti-CD4 antibody [EPR6855] ab133616), anti-CD8 alpha (Anti-CD8 alpha antibody [SP16] ab101500), anti-CD20 (Anti-CD20 antibody [L26] ab9475), anti-CD68 (Anti-CD68 antibody [SP251] ab192847), anti-Cytokeratin 19 (ab52625). Nuclear counter stain shown in dark blue.

  The section was incubated in nine rounds of staining; in the order of Anti-FOXP3 antibody [EPR22102-37] ab215206 (1/100 dilution), Anti-PD1 antibody [NAT105] ab52587 (1/200 dilution), Anti-CD163 antibody [EPR19518] ab182422 (1/300 dilution), Anti-HLA-DR antibody [EPR3692] ab92511 (1/200 dilution), Anti-CD4 antibody [EPR6855] ab133616 (1/600 dilution), Anti-CD8 alpha antibody [SP16] ab101500 (1/300 dilution), Anti-CD20 antibody [L26] ab9475 (1/100 dilution), Anti-CD68 antibody [SP251] ab192847 (1/300 dilution), ab52625 (1/400 dilution); each using a separate fluorescent tyramide signal amplification system.

  Sodium citrate antigen retrieval (pH6.0) was used in between rounds of tyramide signal amplification to remove the antibody from the previous round, to avoid any cross-reactivity.

  Image acquisition was performed with TissueFAXS Spectra (TissueGnostics).

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  Multiplex immunohistochemistry - Anti-Cytokeratin 19 antibody [EP1580Y] - Cytoskeleton Marker (ab52625)

  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 (Anti-Cytokeratin 19 antibody [EP1580Y] - BSA and Azide free 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), Anti-Cytokeratin 19 antibody [EP1580Y] - BSA and Azide free 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.
  
  This data was developed using Anti-Cytokeratin 19 antibody [EP1580Y] - BSA and Azide free ab195872, the same antibody clone in a different buffer formulation. 

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  Immunocytochemistry/ Immunofluorescence - Anti-Cytokeratin 19 antibody [EP1580Y] - Cytoskeleton Marker (ab52625)

  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 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.

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  Flow Cytometry (Intracellular) - Anti-Cytokeratin 19 antibody [EP1580Y] - Cytoskeleton Marker (ab52625)

  Flow cytometry overlay histogram showing left MCF7 positive cells and right negative SH-SY5Y stained with 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 (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.