The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/1000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Use a concentration of 10 µg/ml. Predicted molecular weight: 13.5 kDa.
Use 1µg for 106 cells. ab18536-Rat monoclonal IgG2b, is suitable for use as an isotype control with this antibody.
Application notesIs unsuitable for ICC/IF.
FunctionVariant histone H2A which replaces conventional H2A in a subset of nucleosomes. Nucleosomes wrap and compact DNA into chromatin, limiting DNA accessibility to the cellular machineries which require DNA as a template. Histones thereby play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling. May be involved in the formation of constitutive heterochromatin. May be required for chromosome segregation during cell division.
Sequence similaritiesBelongs to the histone H2A family.
Post-translational modificationsMonoubiquitination of Lys-122 gives a specific tag for epigenetic transcriptional repression. Acetylated on Lys-5, Lys-8 and Lys-12 during interphase. Acetylation disappears at mitosis. Monomethylated on Lys-5 and Lys-8 by SETD6. SETD6 predominantly methylates Lys-8, lys-5 being a possible secondary site. Not phosphorylated.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Histone H2A.Z antibody [8H8] (ab80153)This image is courtesy of an Abreview submitted by Antibody Solutions Ltd.
ab80153 (1/1000) staining Histone H2A.Z in paraffin-embedded Human pancreatic cancer tissue. Tissue underwent fixation in formaldehyde, peroxidase blocking, protein blocking and heat mediated antigen retrieval. After the primary antibody incubation a rabbit anti-rat link was used before the application of the secondary antibody which was goat anti rabbit/mouse conjugated to HRP. For further experimental details please refer to abreview.
Overlay histogram showing HeLa cells stained with ab80153 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab80153, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rat IgG (H+L) (ab98386) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rat IgG2b [RTK4530] ab18541, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
References for Anti-Histone H2A.Z antibody [8H8] (ab80153)
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