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Our Abpromise guarantee covers the use of ab2001 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||Use a concentration of 1 µg/ml.|
|ICC/IF||Use a concentration of 10 µg/ml.|
|WB||1/1000 - 1/2000. Detects a band of approximately 141 kDa.Can be blocked with APAF1 peptide (ab7873).|
ab2001 staining APAF1 in rat intestinal epithelial cells (RIE-1) by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.2% Triton X-100 and blocked with 1.5% serum for 60 minutes at 25°C. Samples were incubated with primary antibody (1/400 with 1x HBSS + 0.02% TritonX-100 + 1.5% FBS) for 3 hour at 25°C. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG polyclonal was used as the secondary antibody at a dilution of 1/2000.
ICC/IF image of ab2001 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 ab2001 at 10µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) 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.
ab2001 staining APAF1 in mouse embryonic fibroblasts by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde and blocked with 10% milk for 1 hour at 25°C. Samples were incubated with primary antibody (1/100 in 1X PBS + 0.02% Tween-20 + 10% milk) for 3 hours at 25°C. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG polyclonal (1/5000) was used as the secondary antibody.
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