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Our Abpromise guarantee covers the use of ab66617 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use a concentration of 1.25 µg/ml. Detects a band of approximately 21 kDa (predicted molecular weight: 21 kDa). Good results were obtained when blocked with 5% non-fat dry milk in 0.05% PBS-T.|
|IHC-P||Use a concentration of 4 - 8 µg/ml.|
|ICC/IF||Use a concentration of 1 µg/ml.|
ICC/IF image of ab66617 stained HeLa 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 (ab66617, 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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung tissue labelling HP1 gamma with ab66617 at 4-8µg/ml. Arrows indiciate positively labelled alveolar cells. Magnification: 400X.