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Our Abpromise guarantee covers the use of ab11826 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.|
|Other||Use at an assay dependent concentration.|
|ELISA||Use at an assay dependent concentration.|
|AP||Use at an assay dependent concentration.|
|IP||Use at an assay dependent concentration. See Fu and Maniatis 1992 reference.|
ab11826 staining SC35 in human fibroblast cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.3% Triton X-100 in PBS and blocked with 5% Normal Goat Serum/0.3% Triton X-100 in PBS for 60 minutes at 25°C. Samples were incubated with primary antibody (1/500 in 1% BSA/ 0.3% Triton X-100 in PBS) for 16 hours at 4°C. An Alexa Flour® 488 goat anti-mouse IgG (H+L), F(ab')2 Fragment Ig was used as the secondary antibody at a dilution of 1/1000.
ab11826 staining cultured human colon adenocarcinoma HT29 cells. Cells were PFA fixed and permeabillized in Triton X100 and Saponin prior to blocking with 1% BSA for 1 hour at RT. The primary antibody was diluted 1/200 and incubated with the sample for 16 hours at 4°C. An Alexa Fluor® 594 conjugated goat anti-mouse IgG3 antibody was used as the secondary.
ICC/IF image of ab11826 stained HeLa 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 ab11826 at 1µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- mouse (ab96879) 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.