Anti-PLK1 antibody [35-206] (ab17056) at 1 µg/ml + A431 cell lysate at 20 µg

Secondary
Rabbit-anti mouse Alexa fluor(680) at 1/10000 dilution

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 67 kDa
Observed band size: 66 kDa why is the actual band size different from the predicted?

Immunofluoresence using ab17056 and either U2OS, HeLaS3 or NIH 3T3 cells.

Plk1 can be seen localising with MKLP in the U2OS cell and with Cdc20 in the HeLaS3 and NIH 3T3 cells shown.

All lanes : Anti-PLK1 antibody [35-206] (ab17056)

Lane 1 : Recombinant PLK1
Lane 2 : U2OS lysate
Lane 3 : HeLaS3

Performed under reducing conditions.

Predicted band size: 67 kDa
Observed band size: 66 kDa why is the actual band size different from the predicted?

Western blot using ab17056.

Lane 1: recombinant PLK1
Lane 2: U2OS lysate
Lane 3: HeLaS3 lysate

15% SDS-PAGE gel

ab17056 at 1/100 staining human kidney tubular epithelial cells by ICC/IF. The cells were formaldehyde fixed, permeabilized with Triton X-100 and blocked with goat serum before incuabtion with the antibody. A goat anti-mouse FITC antibody was used as the secondary.

See Abreview

Overlay histogram showing HCT116 cells stained with ab17056 (red line). The cells were fixed with 4% paraformaldehyde (10 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 (ab17056, 1µg/1x10⁶ cells) 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 IgG2b [PLPV219] (ab91366, 2µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HCT116 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.