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

• ICC/IF

Supplier Data

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 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 (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).

• ICC/IF

Supplier Data

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 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).

• Flow Cyt (Intra)

Supplier Data

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.

• Flow Cyt (Intra)

AbReview46886****

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.

This image is courtesy of an anonymous Abreview

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Alexa Fluor® 488 Anti-Cytokeratin 14 antibody [EP1612Y] (AB192055)

Immunohistochemical analysis of paraffin-embedded Human skin tissue labelling Anti-Cytokeratin 14 with ab192055 at 1/50 (10.0 ug/ml) dilution.

Positive staining on Human skin.

The primary antibody was incubated for 60 mins at room temperature (shown in green). Nuclear DNA was labelled with DAPI (shown in blue). The section was then mounted using Fluoromount®.

The immunostaining was performed on a Leica Biosystems BOND RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 40 mins.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Alexa Fluor® 488 Anti-Cytokeratin 14 antibody [EP1612Y] (AB192055)

Immunohistochemical analysis of paraffin-embedded Human skeletal muscle tissue labelling Anti-Cytokeratin 14 with ab192055 at 1/50 (10.0 ug/ml) dilution.

Negative control : no staining on Human skeletal muscle.

The primary antibody was incubated for 60 mins at room temperature (shown in green). Nuclear DNA was labelled with DAPI (shown in blue). The section was then mounted using Fluoromount®.

The immunostaining was performed on a Leica Biosystems BOND RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 40 mins.