Anti-Cytokeratin 8 antibody [M20] - Cytoskeleton Marker (ab9023)

ab9023 staining Cytokeratin 8 in Human MCF-7 cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.25% Triton X-100 and blocked with 5% BSA for 1 hour at 25°C. Samples were incubated with primary antibody (1/100) at 4°C. An Alexa Fluor® 488-conjugated Goat anti-mouse IgG polyclonal was used as the secondary antibody (1/1000).

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ab9023 staining MCF-7 breast cancer cells, grown in 8 well chamber slides, by ICC/IF.  Cells were fixed with Acetone for 20 minutes at -20°C and permabilized with 0.1% Triton X-100 for 5 minutes at 4°C prior to blocking in 1% fish gelatin for 1 hour at RT.  The primary antibody was diluted 1/100 and incubated with the sample for 1 hour.  An Alexa Fluor® 488 conjugated donkey anti-mouse antibody diluted 1/1000 was used as the secondary.  Slides were mounted using mounting medium containing DAPI.

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The image shows different concentration of total protein from normal and tumour colon tissues.

This picture was kindly supplied as part of the review submitted by Semona Rupchand.

The image shows different concentration of total protein from normal and tumour colon tissues.

This picture was kindly supplied as part of the review submitted by Semona Rupchand.

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Overlay histogram showing MCF-7 cells stained with ab9023 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.5% 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 (ab9023, 2µg/1x10⁶ cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H&L) (ab96879) at 1/250 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. This antibody gave a positive signal in MCF-7 cells fixed with methanol (5 min)/permeabilized in 0.5% PBS-Tween used under the same conditions.

Ab9023 staining cells from a human pancreatic cancer cell line by ICC/IF.  The cells were PFA fixed and permeabilized in 0.1% Triton X-100 prior to blocking in 1% BSA for 30 minutes at 22°C.  Ab9023 was diluted 1/100 and incubated with the sample for 1 hour at 22°C.  An Alexa Fluor® 488 conjugated donkey anti-mouse antibody, diluted 1/1000, was used as the secondary.

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