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Our Abpromise guarantee covers the use of ab21176 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 10 µg/ml.|
|IHC-Fr||1/1000. PubMed: 18219389|
|WB||1/1000 - 1/2000.|
ab21176 at 10 - 20 µg/mL staining Luciferase in transfected HEK293 cells by ICC/IF. The cells were fixed with methanol and acetone. A FITC conjugated Anti-Rabbit IgG was used as the secondary antibody.
ab21176 at 1/200 dilution staining engineered adult rat stromal stem cells by ICC/IF. The cells were paraformaldehyde (2%) fixed and Triton X-100 (0.1%) was used for cell permealization (15 min incubation time). The cells were incubated with the antibody overnight at 4C. The image shows Luciferase (green-upper right panel), counterstained cell nuclei (DAPI-blue-upper left panel), overlay (lower left panel) and a phase contrast image (lower right panel). The image was taken with a confocal laser scanning microscope equipped with an additional laser differential Interference Contrast (DIC) mode.
ab21176 staining Firefly Luciferase in Mouse cerebellum tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with acetone, permeabilized with PBS + 0.1% Triton X-100 (PBST) for 10 minutes and blocked with 10% serum for 1 hour at 22°C. Samples were incubated with primary antibody (1/1500 in 10% Goat serum in PBST) for 1 hour at 22°C. An Alexa Fluor® 555-conjugated Goat anti-rabbit IgG polyclonal (1/1000) was used as the secondary antibody.
ab21176 staining mouse mammary carcinoma cells by ICC/IF. Cells were PFA fixed and permeabilized in 0.1% Triton X-100 prior to blocking with a commercial blocking agent. The primary antibody was diluted 1/100 and incubated with the sample for 1 hour at 25°C. An Alexa Fluor® 555 conjugated goat anti-rabbit antibody was used as the secondary. The image shows firefly luciferase (red) in mouse tumour cells and cell nuclei counterstained with Hoechst (blue).
Immunocytochemical analysis analysis of HEK-293T cells overexpressing luciferase labelling Luciferase with ab21176 at a concentration of 10μg/mL. The secondary antibody was a Goat Anti-Rabbit IgG, FITC conjugate.
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