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Synthetic peptide conjugated to KLH derived from within residues 1200 - 1300 of Human Ki67.
(Peptide available as ab15581.)
Our Abpromise guarantee covers the use of ab15580 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC - Wholemount||Use at an assay dependent concentration.|
|IHC-P||Use a concentration of 0.1 - 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|IHC-FrFl||1/500. (see Abreview)|
|IHC-Fr||1/100 - 1/1000.|
|ICC/IF||1/100 - 1/1000.|
|ICC||Use at an assay dependent concentration.|
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 345, 395 kDa (predicted molecular weight: 359 kDa).Can be blocked with Ki67 peptide (ab15581).|
|IHC-FoFr||Use at an assay dependent concentration.|
Fluorescent confocal microscopy (20x) of mouse (P0) olfactory bulb, outer glomeruli layer, showing Ki67 immunoreactivity (ab15580; 1/1000; overnight at RT, 0.25% TX-100 no blocking step) using a secondary goat anti-rabbit fluorescent antibody (Alexa Fluor 488;1/300 2h at RT.
SK-N-SH cells were permitted to grow to confluency, then serum starved for 48 hours and predominantly driven into G0. The cells were then paraformaldehyde fixed and immunofluorescently labelled with anti-Ki67 (ab15580) at a dilution of 1/1000. The majority of the cells show little or no Ki67 staining, indicating they are in G0 arrest (red cells). Two cells however show strong nucleolar Ki67 staining indicating they are still cycling (green cells). The DNA is stained with DAPI and is shown in red. The Ki67 staining is shown in green. x 63 magnification.
Similar results were seen with an asynchronous population of HeLa cells. The Ki67 staining was localised to the periphery of the nucleoli and throughout the nucleoplasm of proliferating cells. (This data is not shown but is available upon request).
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