Our Abpromise guarantee covers the use of ab10543 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-Fr||Use at an assay dependent concentration.|
|ICC/IF||Use a concentration of 2.5 µg/ml.|
|WB||Use a concentration of 0.5 - 1 µg/ml. Detects a band of approximately 15 kDa.
We advise you to centrifuge this product vial before use.
|ICC||Use at an assay dependent concentration.|
|Flow Cyt||Use at an assay dependent concentration.|
ab10543 staining Histone H3 (phospho S28) in Human neural progenitor cells from iPS cell line by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.1% Triton and blocked with 1% serum, 0.1% BSA in PBS for 30 minutes at room temperature. Samples were incubated with primary antibody (2ug/ml in blocking buffer) for 16 hours at 4°C. An Alexa Fluor® 488-conjugated Goat anti-rat IgG2a polyclonal was used as the secondary antibody (1/500). Total cells were stained using DAPI (blue)
This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab10543 overnight at 4°C. Antibody binding was detected using an anti-mouse antibody conjugated to HRP, and visualised using ECL development solution.
ab10543 staining Histone H3 (phospho S28) in Mouse neural stem cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde and blocked with 4% serum for 1 hour at 20°C. Samples were incubated with primary antibody (1/500 in PBS + 4% serum) for 16 hours at 4°C. An Alexa Fluor® 546-conjugated anti-rat polyclonal was used as the secondary antibody (1/500). DAPI is stainded blue
ab10543 staining HeLa cells by ICC/IF. The cells were fixed with paraformaldehyde and permeabilized with 0.5% Triton X100 in PBS. The cells were then stained with ab10543 at 1/1000 in PBS for 1h at 22°C. A goat anti-rat Alexa Fluro 488 (ab150157) at 1/200 was used as the secondary antibody. Nuclei are stained in red with DAPI. The antibody produces the expected mitotic-associated staining pattern and is extremely strong. MeOH fixed samples were also evaluated and produced a similar, strong staining pattern in mitotic cells.
ab10543 staining Histone H3 (phospho S28) in mouse embryonic brain tissue section by Immunohistochemistry (Frozen sections). Tissue samples were fixed with paraformaldehyde and blocking with 10% serum for 1 hour at 250C was performed. The sample was incubated with primary antibody (1/300) at 40C for 16 hours. An Alexa Fluor®488-conjugated Goat polyclonal to rat IgG was used as secondary antibody at 1/500 dilution.