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Our Abpromise guarantee covers the use of ab15481 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 1 µg/ml.|
|IHC-P||1/100. Antigen retrieval is not essential but may optimise staining.
10min at RT.
Immunohistochemistry (Formalin-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue labelling MUC1 with ab15481.
ab15481 at 1/200 staining mouse mammary gland tissue sections by IHC-P. The tissue was paraformaldehyde fixed and blocked with BSA. A heat mediated antigen retrieval step was performed. The antibody was incubated with the tissue for 16 hours and then an Alexa-Fluor 488 conjugated goat anti-rabbit antibody was used as the secondary. MUC1 staining (luminal cells) is shown in green. The nuclei were counterstained with DAPI and staining is shown in blue.
ICC/IF image of ab15481 stained Hek293 cells. The cells were 100% methanol fixed (5 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 (ab15481, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 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.
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