Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [IGX4228R-1] to Histone H2B
- Suitable for: WB, ChIP
- Reacts with: Mouse, Human, Recombinant fragment
Product nameAnti-Histone H2B antibody [IGX4228R-1]
See all Histone H2B primary antibodies
DescriptionRabbit monoclonal [IGX4228R-1] to Histone H2B
SpecificityThis antibody is a recombinant chimeric antibody. Rabbit chimeric monoclonal antibody (Human Fab/ Rabbit Fc).
Tested applicationsSuitable for: WB, ChIPmore details
Species reactivityReacts with: Mouse, Human, Recombinant fragment
Synthetic peptide corresponding to Human Histone H2B.
- WB: CTH, HeLa whole cell, HeLa Nuclear, NIH3T3 whole cell, NIH3T3 Nuclear and Histone H2B Recombinant Protein lysates. ChIP: HeLa and NIH3T3.
This antibody is a recombinant chimeric antibody. Rabbit chimeric monoclonal antibody (Human Fab/ Rabbit Fc).
For recommended secondary antibodies -
Goat Anti-Rabbit IgG H&L (HRP) - ab205718
Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) - ab150077
Goat Anti-Rabbit IgG H&L (Alexa Fluor® 555) - ab150078
Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) - ab150079
Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) - ab150080
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 bufferConstituents: 0.484% Tris buffer, 0.45% Glycine, 40% Glycerol, 1.044% Sodium chloride
Concentration information loading...
PurityImmunogen affinity purified
Our Abpromise guarantee covers the use of ab213225 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use a concentration of 0.25 µg/ml. Detects a band of approximately 17 kDa (predicted molecular weight: 14 kDa).|
|ChIP||Use 2 µg for 25 µg of chromatin.|
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 antibody [IGX4228R-1] (ab213225) at 0.25 µg/ml
Lane 1 : CTH (Calf Thymus Histone) at 0.5 µg
Lane 2 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 20 µg
Lane 3 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate at 10 µg
Lane 4 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate at 20 µg
Lane 5 : NIH 3T3 (Mouse embryonic fibroblast cell line) Nuclear Lysate at 10 µg
Lane 6 : Histone H2B Recombinant Protein at 0.1 µg
Lane 7 : Histone H3 Recombinant Protein at 0.1 µg
All lanes : Peroxidase AffiniPure Goat Anti-Rabbit IgG (H+L) at 1/50000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 14 kDa
Observed band size: 17 kDa why is the actual band size different from the predicted?
Exposure time: 1 minute
This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before being incubated with ab213225 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.
Chromatin was prepared from HeLa cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 2µg of ab213225 (blue), and 20µl of Protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Taqman approach). Primers and probes are located in the first kb of the transcribed region.
ab213225 has not yet been referenced specifically in any publications.