Recombinant full length protein corresponding to Human PLK1. His-PLK1 full length purified from Sf9 cells.
Database link: P53350
Our Abpromise guarantee covers the use of ab17057 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Indirect ELISA||Use at an assay dependent concentration.|
|IP||Use at an assay dependent concentration.|
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 66 kDa (predicted molecular weight: 68 kDa).|
|ICC/IF||1/200. PubMed: 19033445|
|Flow Cyt||Use 1µg for 106 cells. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.|
10% SDS-PAGE gel.
Immunofluoresence using ab17057 and either HeLa S3, NIH/3T3 (Mouse embyro fibroblast cell line) or U-2 OS (Human bone osteosarcoma epithelial cell line) cells.
In panel one HeLa (Human epithelial cell line from cervix adenocarcinoma) cells were stained with ab17057 (green) and DAPI. In the second panel, cells were stained with ab17057 (green) and SH-CREST (red), which stains the centromeres. Fix 30 minutes on ice in 4% formaldehyde in PEM. Quench autofluorescence 2 x 5 minutes with 1 mg/ml Na borohydride or 100 mM ammonium chloride in PEM. Permeabilized for 30 minutes with 0.5% TX-100 in PEM. Block 30 minutes in 5% milk in TBST. Primary antibody incubated overnight at 4°C diluted 1/400 in 5% milk in TBST. Secondary antibody incubated 1 hour at RT diluted in 5% milk in TBST. Post-fix 20 minutes on ice in 4% formaldehyde in PEM. Quench autofluorescence 2 x 5 minutes with ammonium chloride in PEM. Counterstain with DAPI in TBST. Mount with ProLong Gold antifade reagent from Invitrogen.
Notes: Ample washing between each step.
TBST = Tris buffered saline + 0.1% Tween. PEM = 80 mM K-PIPES, pH 6.8, 5 mM EGTA, 2 mM MgCl2.
Overlay histogram showing U-2 0S (Human bone osteosarcoma epithelial cell line) cells stained with ab17057 (red line). The cells were fixed with 80% methanol (10 minutes) and then permeabilized with 0.1% PBS-Triton X-100 for 15 minutes. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab17057, 1 µg/1 x 106 cells) for 30 minutes at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H+L) (ab150117) at 1/2000 dilution for 30 minutes at 22°C.
Isotype control antibody (black line) was mouse IgG1 [15-6E10A7] (ab170190, 1 µg/1 x 106 cells) used under the same conditions.
Acquisition of >5,000 events were collected using a 50 mW Blue laser (488nm) and 530/30 bandpass filter.