Anti-pan Cytokeratin antibody [5D3 + LP34] (ab17153)

  Immunocytochemistry/ Immunofluorescence-pan Cytokeratin antibody [5D3 + LP34](ab17153)

ICC/IF image of ab17153 stained HeLa 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 (ab17153, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 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.

  Western blot - pan Cytokeratin antibody [5D3 + LP34] (ab17153)

Anti-pan Cytokeratin antibody [5D3 + LP34] (ab17153) at 1/500 dilution ((in 5% Marvel TBS-T). Incubation for 1 hour at 22°C) + Whole cell lysate of human HT1080 cells at 25 µg

Secondary
An HRP-conjugated Sheep anti-mouse IgG monoclonal at 1/5000 dilution
developed using the ECL technique

Performed under reducing conditions.

Observed band size : 45 kDa (why is the actual band size different from the predicted?)


Exposure time : 15 minutes

Blocking Step: 5% Milk for 1 hour at 22°C

This image is courtesy of an anonymous Abreview

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