Our Abpromise guarantee covers the use of ab18262 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ChIP||Use at an assay dependent dilution.|
|ICC/IF||Use at an assay dependent dilution.|
|WB||Use at an assay dependent dilution. Detects a band of approximately 14 kDa (predicted molecular weight: 13 kDa).|
ChIP analysis of Acetyl-H2A.Z at the p21 transcription start site with or without the HDAC inhibitor TSA (50ng/ml). Chromatin was prepared from MDA-MB231 cells. Cells were fixed with formaldehyde for 10min. Samples were sonicated to generate DNA fragments between 500 and 700 bp. The ChIP was performed with 50µg of chromatin, fragments were immunoprecipitated using 2µg of antibody ab18262, and an HA antibody was used as control. The precipitated DNA was amplified by real-time PCR, with primer sets designed to amplify the promoter and the coding region of the p21 gene.
This image is courtesy of an anonymous Abreview
ab18262 staining Histone H2A.Z (acetyl K4 + K7 + K11) in HeLa cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.5% Triton X-100 and blocked with 3% BSA for 1 hour at 22°C. Samples were incubated with primary antibody (1/500 in PBS + 1% BSA) for 1 hour at 22°C. An Alexa Fluor® 488-conjugated donkey anti-sheep IgG polyclonal (1/1000) was used as the secondary antibody.
This image is courtesy of Prof Colyn Crane-Robinson
This image is courtesy of Prof Colyn Crane-RobinsonNucleosomes from 10-day chicken brain tissue were used in ChIP experiments. An acetic acid / urea / Triton gel was run and the corresponding western blot using ab18262 was used to compare proteins in the Bound chromatin fraction with those of the Input and Unbound fractions. In lane 8 there is a strong diffuse band corresponding to a hyper-acetylated H2A.Z.