Key features and details
- Rabbit polyclonal to Histone H2B (butyryl K5)
- Suitable for: ChIP, WB, IP, ICC/IF
- Reacts with: Human
- Isotype: IgG
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.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
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
Constituents: 50% Glycerol (glycerin, glycerine), PBS, 0.03% Proclin 300
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.
- GL105 antibody
- H2B GL105 antibody
- H2B histone family member O antibody
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.
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
ab235429 has not yet been referenced specifically in any publications.