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Thioglycollate stimulated peritoneal macrophages from C57/BL mice
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Our Abpromise guarantee covers the use of ab90247 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use a concentration of 5 - 10 µg/ml.|
|WB||Use a concentration of 10 µg/ml. Detects a band of approximately 160 kDa (predicted molecular weight: 98 kDa).|
Ab90247 stained Raw 264.7 cells. The cells were 100% methanol fixed (5 minutes) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Triton for 1hour at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab90247 at 10µg/ml) overnight at +4°C. The secondary antibody (pseudo-colored green) was Alexa Fluor® 488 goat anti-rat IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1hour at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.