Anti-pan Cytokeratin antibody [C-11] (ab7753)

Fluorescence multiplex immunohistochemical analysis of normal human tonsil tissue (formalin-fixed paraffin-embedded section).

Merged staining of anti-PD1 (ab237728; orange; Opal™520), anti-PDL1 (ab237726; green; Opal™540), anti-CD68 (ab192847; yellow; Opal™570), anti-CD3 (ab16669; red; Opal™620), anti-Ki67 (ab16667; light blue; Opal™650) and anti-PanCK (ab7753; grey; Opal™690).

The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 7-color automation IHC kit (NEL821001KT, Akoya Biosciences®).

The section was incubated in six rounds of staining; in the order of ab237728 (1/500 dilution), ab237726 (1/500 dilution), ab192847 (1/300 dilution), ab16669 (1/300 dilution), ab16667 (1/200 dilution) and ab7753 (1/200 dilution); each using a separate fluorescent tyramide signal amplification system.

Sodium citrate antigen retrieval (Leica ER1, pH6.0, 30 minutes) was used in between rounds of tyramide signal amplification to remove the antibody from the previous round, to avoid any cross-reactivity.

DAPI (dark blue) was used as a nuclear counter stain.

Microscopy and pseudocoloring of individual Opal™ dyes was performed using a Vectra Polaris.

ab7753 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 ab7753 at 1/250 in TBS/BSA/azide for 2h at 21°C. A biotinylated goat anti-rabbit polyclonal antibody at 1/200 was used as the secondary antibody.

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ab7753 staining pan Cytokeratin in Rat urinary bladder tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with formaldehyde and blocked with 1% BSA for 10 minutes at 21°C. Samples were incubated with primary antibody (1/150 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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ab7753 staining pan Cytokeration in the Human NSCLC cell line by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.3% Triton and blocked with 5% serum for 1 hour at room temperature. Samples were incubated with primary antibody (1/500 in 0.3% Triton, 5% Goat Serum in PBS) for 1 hour. An Alexa Fluor® 555-conjugated Goat polyclonal (1/500) was used as the secondary antibody.

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ab7753 staining rat embryonic skin/organ 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 ab77539 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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ab7753 staining pan Cytokeration in Cow Airway Epithelial cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with formaldehyde, permeabilized with 0.5% Triton X-100 and blocked with 1% serum for 15 minutes at 37°C. Samples were incubated with primary antibody (1/500 in PBS/1%BSA) for 1 hour at 37°C. A Biotin-conjugated Goat anti-mouse IgG polyclonal (1/50) was used as the secondary antibody.

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ab7753 staining pan Cytokeratin in Mouse liver 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 a 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 (1/250) was used as the secondary antibody.

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ab7753 staining pan Cytoketatin in mouse hepatocytes by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde and blocked with 0.5% BSA for 1 hour at 23°C. Samples were incubated with primary antibody (1/500) for 16 hours at 4°C. A Cy3^®-conjugated goat anti-mouse IgG polyclonal (1/1000) was used as the secondary antibody.

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ab7753 staining pan Cytokeratin in guinae pig breast carcinoma tissue section by Immunohistochemistry (Formalin/PFA fixed paraffin-embedded sections).