Product nameAnti-Histone H2B (butyryl K5) antibody
See all Histone H2B primary antibodies
DescriptionRabbit polyclonal to Histone H2B (butyryl K5)
Tested applicationsSuitable for: ChIP, WB, IP, ICC/IFmore details
Species reactivityReacts with: Human
Predicted to work with: Rat
Synthetic peptide corresponding to Human Histone H2B (butyryl K5).
Database link: P62807
- ChIP: Chromatin prepared from HeLa cells treated with 30mM sodium butyrate for 4 hours. IP: HEK-293 whole cell lysate. ICC/IF: HeLa cells (treated with 30mM sodium butyrate for 4 hours). WB: HeLa, HEK-293, A549 and HepG2 whole cell lysates, treated with 30 mM sodium butyrate for 4 hours.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage bufferpH: 7.40
Preservative: 0.03% Proclin
Constituents: 50% Glycerol, PBS
Concentration information loading...
PurityImmunogen affinity purified
Our Abpromise guarantee covers the use of ab235429 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ChIP||Use 5µg for 106 cells.|
|WB||1/500 - 1/5000. Predicted molecular weight: 14 kDa.|
|IP||1/200 - 1/2000.|
|ICC/IF||1/10 - 1/100.|
RelevanceCore 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. Subunit structure The nucleosome is a histone octamer containing two molecules each of H2A, H2B, H3 and H4 assembled in one H3-H4 heterotetramer and two H2A-H2B heterodimers. The octamer wraps approximately 147 bp of DNA. Post-translational modification Monoubiquitination at Lys-35 (H2BK34Ub) by the MSL1/MSL2 dimer is required for histone H3 'Lys-4' (H3K4me) and 'Lys-79' (H3K79me) methylation and transcription activation at specific gene loci, such as HOXA9 and MEIS1 loci. Similarly, monoubiquitination at Lys-121 (H2BK120Ub) 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. H2BK120Ub also acts as a regulator of mRNA splicing: deubiquitination by USP49 is required for efficient cotranscriptional splicing of a large set of exons. Phosphorylation at Ser-37 (H2BS36ph) by AMPK in response to stress promotes transcription. Phosphorylated on Ser-15 (H2BS14ph) 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. GlcNAcylation at Ser-113 promotes monoubiquitination of Lys-121. It fluctuates in response to extracellular glucose, and associates with transcribed genes. Crotonylation (Kcr) is specifically present in male germ cells and marks testis-specific genes in post-meiotic cells, including X-linked genes that escape sex chromosome inactivation in haploid cells. Crotonylation marks active promoters and enhancers and confers resistance to transcriptional repressors. It is also associated with post-meiotically activated genes on autosomes.
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All lanes : Anti-Histone H2B (butyryl K5) antibody (ab235429) at 1/100 dilution
Lane 1 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate, treated (+) with 30mM sodium butyrate for 4 hours
Lane 2 : HEK-293 (human epithelial cell line from embryonic kidney) whole cell lysate, treated (+) with 30mM sodium butyrate for 4 hours
Lane 3 : A549 (human lung carcinoma cell line) whole cell lysate, treated (+) with 30mM sodium butyrate for 4 hours
Lane 4 : HepG2 (human liver hepatocellular carcinoma cell line) whole cell lysate, treated (+) with 30mM sodium butyrate for 4 hours
Lane 5 : HeLa whole cell lysate, untreated
Lane 6 : HEK-293 whole cell lysate, untreated
Lane 7 : A549 whole cell lysate, untreated
Lane 8 : HepG2 whole cell lysate, untreated
All lanes : Goat polyclonal to rabbit IgG at 1/50000 dilution
Predicted band size: 14 kDa
HeLa (human epithelial cell line from cervix adenocarcinoma) cells (treated with 30 mM sodium butyrate for 4 hours) labeling Histone H2B (butyryl K5) with ab235429 at 1/15 dilution in ICC/IF.
The cells were fixed in 4% formaldehyde, permeabilized using 0.2% Triton X-100 and blocked in 10% normal goat serum. The cells were then incubated with the antibody overnight at 4°C. The secondary antibody was Alexa-Fluor® 488-conjugated Goat Anti-Rabbit IgG (H+L).
Histone H2B (butyryl K5) was immunoprecipitated from HEK-293 (human epithelial cell line from embryonic kidney) whole cell lysate using 5 µg of ab235429.
Lane 1: Rabbit control IgG in HEK-293 whole cell lysate.
Lane 2: ab235429 IP in HEK-293 whole cell lysate.
Lane 3: HEK-293 whole cell lysate (20 µg input).
For western blotting an HRP-conjugated Protein G antibody was used as the secondary antibody at 1/2000 dilution.
HeLa (human epithelial cell line from cervix adenocarcinoma) (106, treated with 30 mM sodium butyrate for 4 hours) were treated with Micrococcal Nuclease, sonicated, and immunoprecipitated with 5 μg ab235429 or a control normal rabbit IgG. The resulting ChIP DNA was quantified using real-time PCR with primers against the β-Globin promoter.
ab235429 has not yet been referenced specifically in any publications.