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Synthetic peptide derived from residues 50 - 150 of Human Skp1.
Our Abpromise guarantee covers the use of ab10546 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/500. Detects a band of approximately 19 kDa (predicted molecular weight: 18.5 kDa).|
|IHC-P||Use at an assay dependent concentration.|
|ICC/IF||Use a concentration of 1 µg/ml.|
ICC/IF image of ab10546 stained MCF7 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 (ab10546, 1µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 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 using ab10546 at 1/500.
Lane 1: Hela nuclear
Lane 2: Hela
Lane 3: HEK293
Lane 4: Hela nuclear + ab10546 blocking peptide (1µg/ml)
Lane 5: Hela + ab10546 blocking peptide (1µg/ml)
Lane 6: HEK293 + ab10546 blocking peptide (1µg/ml)
Secondary ab: Goat anti-rabbit IgG ab6721 (1/5000)
Exposure time: 2 mins.
ab10546 was used in immunohistochemistry with paraffin embedded sections of human tonsil, using DAB as a chromogen (brown). Counterstaining of nuclei was performed with haemotoxylin (blue).
Strong staining of epithelial cell nuclei is seen (and some staining of the cytoplasm) in this image.