The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/250 - 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
1/2000. Detects a band of approximately 15 kDa (predicted molecular weight: 14 kDa).
1/250 - 1/500.
Application notesIs unsuitable for IP.
FunctionCore component of nucleosome. 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.
Sequence similaritiesBelongs to the histone H2B family.
Post-translational modificationsMonoubiquitination of Lys-121 by the RNF20/40 complex gives a specific tag for epigenetic transcriptional activation and is also prerequisite for histone H3 'Lys-4' and 'Lys-79' methylation. It also functions cooperatively with the FACT dimer to stimulate elongation by RNA polymerase II. Phosphorylated on Ser-15 by STK4/MST1 during apoptosis; which facilitates apoptotic chromatin condensation. Also phosphorylated on Ser-15 in response to DNA double strand breaks (DSBs), and in correlation with somatic hypermutation and immunoglobulin class-switch recombination.
IHC image of ab40883 staining Histone H2B (acetyl K12) in human colon formalin fixed paraffin embedded tissue sections, performed on a Leica Bond. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab40883, 1/200 dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. No primary antibody was used in the negative control (shown on the inset).
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
Western blot - Histone H2B antibody [EP858Y] (ab40883)
Anti-Histone H2B (acetyl K12) antibody [EP858Y] (ab40883) at 1/2000 dilution + C6+TSA cell lysate at 10 µg
Immunohistochemical analysis of paraffin-embedded human colon carcinoma, using anti-Histone H2B ab40883 at a 1:250 dilution.Immunohistochemical analysis of paraffin-embedded human colon carcinoma, using anti-Histone H2B ab40883 at a 1:250 dilution.
ab40883 (1:250) staining of HeLa cells by immunofluorescence.
Immunocytochemistry/ Immunofluorescence - Anti-Histone H2B (acetyl K12) antibody [EP858Y] (ab40883)Image courtesy of an abreview submitted by Dr. Kirk McManus, Univ. of Manitoba/Cancer Care MICB, Canada
ab40883 (1/500) staining Histone H2B (acetyl K12) in assynchronous HeLa cells (green). Cells were fixed with paraformaldehyde, permeabilised using 0.5% Triton X100 and counterstained with DAPI in order to highlight the nucleus (red). For further experimental details please refer to Abreview.