ab86734 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab86734 at 1/100 dilution overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- mouse (ab96879) IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in methanol fixed (100%, 5min) HeLa, Hek293. HepG2, and MCF-7 cells, also in formaldehyde fixed (4%, 10min) HeLa, Hek293, and MCF-7 cells at 1/100 dilution.

Paraffin embedded, 0.1% Triton X-100 permeabilized mouse skin stained for pan Cytokeratin with ab86734 at a 1/100 dilution. Heat mediated - Buffer/Enzyme Used: Trypsin enzyme for 15 minutes at RT. 10% serum used to block for 1 hour at RT. Primary incubation for 16 hours at 4°C. Secondary: Goat polyclonal conjugated to biotin used at a 1/100 dilution.

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ab86734 staining pan Cytokeratin in rat squamous epithelial tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).
Tissue was fixed in formaldehyde and an enzymatic antigen retrieval step was performed using Proteinase K. Samples were then blocked using 1% BSA for 25 minutes at 25°C, followed by incubation with ab86734 at a 1/100 dilution for 16 hours at 25°C. The secondary used was an undiluted HRP goat polyclonal.

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Overlay histogram showing A431 cells stained with ab86734 (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 (ab86734, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.

ab86734 staining pan Cytokeratin in pig small intestine tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 1% BSA for 10 minutes at 21°C; antigen retrieval was by heat mediation in citric acid. Samples were incubated with primary antibody (1/250 in TBS/BSA/azide) for 2 hours at 21°C. A Biotin-conjugated goat anti-mouse IgG polyclonal (1/200) was used as the secondary antibody.

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Formaldehyde fixed dog lung tissue stained for pan Cytokeratin with ab86734 at a 1/250 dilution. Heat mediated - Buffer/Enzyme Used: Citric acid. 1% BSA used for blocking for 10 minutes at RT. Primary incubation for 2 hours at RT in TBS/BSA/Azide. Secondary: Goat polyclonal conjugated to biotin used at a 1/200 dilution.

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Formaldehyde fixed goat lung tissue stained for pan Cytokeratin with ab86734 at a 1/250 dilution. Heat mediated - Buffer/Enzyme Used: Citric acid. 1% BSA used for blocking for 10 minutes at RT. Primary incubation for 2 hours at RT in TBS/BSA/Azide. Secondary: Goat polyclonal conjugated to biotin used at a 1/200 dilution.

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Formaldehyde fixed chicken lung tissue stained for pan Cytokeratin with ab86734 at a 1/200 dilution. Heat mediated - Buffer/Enzyme Used: Citric acid. 1% BSA used for blocking for 10 minutes at RT. Primary incubation for 16 hours at RT in TBS/BSA/Azide. Secondary: Goat polyclonal conjugated to biotin used at a 1/200 dilution.

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Skin stained for pan Cytokeratin with ab86734 at a 1/100 dilution.

ab8673 staining human skin sections by IHC-P. The tissue was fixed with formaldehyde and a heat mediated antigen retrival step was performed with citric acid pH 6. Blocking of the sample was done with 1% BSA for 10 minutes at 21°C, followed by staining with ab8673 at 1/250 in TBS/BSA/azide for 2h at 21°C. A biotinylated goat anti-mouse polyclonal antibody at 1/200 was used as the secondary antibody.

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