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Our Abpromise guarantee covers the use of ab3642 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||1/25. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|IHC-Fr||Use at an assay dependent concentration.|
|WB||Use a concentration of 1 - 2 µg/ml. Detects a band of approximately 14 kDa (predicted molecular weight: 14 kDa).|
|ICC/IF||Use a concentration of 1 µg/ml.|
Confocal image showing nuclear staining increased after HeLa cells transfected with CDKN2A/p14ARF.
ICC/IF image of ab3642 stained HeLa cells. The cells were 4% paraformaldehyde fixed and then incubated in 0.1% trixtonX-100 to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with primary antibody ab3642 at a dilution of 1/250. An AlexaFluor®594 goat anti-mouse secondary IgG (ab150077) was used at a 1/1000 dilution. Anti-tubulin (ab7291) and an AlexaFluor®594 goat anti-mouse IgG (ab150120) were used as counterstains, both at a dilution of 1/1000. DAPI was used to stain the cell nuclei blue.
ICC/IF image of ab3642 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 ab3642 at 1µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) 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.
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