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Our Abpromise guarantee covers the use of ab833 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||1/400. PubMed: 18635871|
|ICC||Use at an assay dependent concentration.|
|IHC-P||1/25 - 1/50. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. We recommend using an ABC system for detection of this antibody.|
|IHC-Fr||1/25 - 1/50. 1/25 - 1/50. We suggest an incubation period of 30 minutes at room temperature and to use DAB to stain the protein.|
Immunocytochemistry/ Immunofluorescence analysis of HeLa cells labeling Ki67 with ab833 at 1/50 dilution.
HeLa cells are the classic example of an immortalized human epithelial cell line from a fatal cervical carcinoma transformed by human papillomavirus 18 (HPV18) that grown as adherent cells layer and maintain contact inhibition in vitro. 2% paraformaldehyde for 15 min was used to fix the cells, and permealized the cells with Triton-X100 (0.1%) for 10 min and then incubated cells with primary antibody Ki-67 (1/50) for overnight at 4 degree centigrade. Image was taken using a confocal laser scanning microscope that equipped with an additional differential Interference Contrast (DIC) mode. Image here demonstrate DAPI (blue-nuclear stain, upper left panel), Ki-67 (Green, upper right panel, shows nuclear localization in actively dividing cells), Hela cells in culture-DIC (phase) image (lower left panel) and superimpose image (lower right panel). The expression of the human Ki-67 protein is strictly associated with cell proliferation. During interphase, the antigen is exclusively detected as several prominent specks within the nucleus and a weak finely dispersed staining throughout the nucleoplasm, whereas in mitosis most of the protein is relocated to the surface of the chromosomes. The fact that the Ki-67 protein is present during all active phases of the cell cycle [G(1), S, G(2), and mitosis], but is absent from resting cells [G(0)], makes it an excellent marker for determining the so-called Ki-67 staining index (Ki67-SI) of a given cell population.
ab833 - immunohistochemistry
Formalin fixed paraffin embedded human tonsil stained with Ki-67, using ABC and AEC chromogen.
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