Alexa Fluor® 647 Anti-Cytokeratin 14 antibody [LL002]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Alexa Fluor® 647 Anti-Cytokeratin 14 antibody [LL002] (AB321783)

Negative immunofluorescence staining of Cytokeratin 14 in a section of formalin-fixed paraffin-embedded human brain*.

Performed on a Leica BOND. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6.0) using retrieval settings of 110°C for 40 minutes. The section was then incubated at room temperature for 1 hour with ab321783 at 1/1000 dilution. Nuclear DNA was labelled with DAPI (shown in blue). The section was then mounted using Dako Fluorescence Mounting Medium®.

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

For other IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antigen retrieval conditions, antibody concentrations and incubation times.

*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-Cytokeratin 14 antibody [LL002] (AB321783)

ab321783 staining Cytokeratin14 in A-431 cells, with negative expression in KRT14 knockout A-431 cells. The cells were fixed with 4% formaldehyde (10 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 ab195887 at 0.5 μg/ml (shown in Magenta). ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 488) (shown in green) was used at 2 μg/ml for structural counterstaining.

Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

This product also work with 100% methanol (5 min) fixation under the same testing conditions.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Alexa Fluor® 647 Anti-Cytokeratin 14 antibody [LL002] (AB321783)

Immunofluorescence staining of Cytokeratin 14 staining in a section of formalin-fixed paraffin-embedded human skin normal*.

Performed on a Leica BOND. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6.0) using retrieval settings of 100°C for 40 minutes. The section was then incubated at room temperature for 1 hour with ab321783 at 1/1000 dilution (shown in magenta). Nuclear DNA was labelled with DAPI (shown in blue). The section was then mounted using Dako Fluorescence Mounting Medium®.

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

For other IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antigen retrieval conditions, antibody concentrations and incubation times.

*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.