Alexa Fluor® 488 Anti-Cytokeratin 14 antibody [EP1612Y]

4 product images

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  This image is courtesy of an anonymous customer review

  Flow Cytometry (Intracellular) - Alexa Fluor® 488 Anti-Cytokeratin 14 antibody [EP1612Y] (ab192055)

  ab192055 staining Cytokeratin 14 in CK14 bright positive and low cell lines by Flow Cytometry. Cells were fixed with paraformaldehyde and permeabilized with PBS + 5% FBS + 2% Triton X-100. The sample was incubated with the primary antibody (1/50 in PBS + 5% FBS + 0.2% Triton X-100) for 45 minutes at 4°C.

  Gating Strategy: Doublet were excluded based on SSC - FSC profile.

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  Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 488 Anti-Cytokeratin 14 antibody [EP1612Y] (ab192055)

  ab192055 staining KRT14 in wild-type A431 cells (top panel) and KRT14 knockout A431 cells (bottom panel). The cells were fixed with 100% methanol (5 min) then 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 ab192055 at 1/100 dilution and Alexa Fluor® 647 Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker ab190573 (Rabbit monoclonal to alpha Tubulin - Alexa Fluor^® 647) at 1/250 dilution 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) - Alexa Fluor® 488 Anti-Cytokeratin 14 antibody [EP1612Y] (ab192055)

  Overlay histogram showing A431 cells stained with ab192055 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton X-100 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 (ab192055, 1/500 dilution) for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) Alexa Fluor® 488 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 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

  This antibody gave a positive signal in A431 fixed with 4% formaldehyde (10 min)/permeabilized with 0.1% PBS-Triton X-100 for 20 min used under the same conditions.

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  Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 488 Anti-Cytokeratin 14 antibody [EP1612Y] (ab192055)

  ab192055 staining Cytokeratin 14 in A431 cells. The cells were fixed with 100% methanol (5min), permeabilized in 0.1% Triton X-100 for 5 minutes and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab192055 at a working dilution of 1 in 100 (shown in green) 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 1h with an Alexa Fluor^® 594 Goat anti-Mouse secondary (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.

  This product also gave a positive signal in 4% formaldehyde (10 min) fixed A431 cells under the same testing conditions.

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