Anti-Histone H2B antibody (ab134211)
- Datasheet
- References (2)
- Protocols
Overview
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Product name
Anti-Histone H2B antibody
See all Histone H2B primary antibodies -
Description
Chicken polyclonal to Histone H2B -
Host species
Chicken -
Tested applications
Suitable for: IHC-P, ICC/IF, WBmore details -
Species reactivity
Reacts with: Human
Predicted to work with: Mouse, Rat, Horse, Cow, Dog, Chimpanzee, Drosophila melanogaster, Cynomolgus monkey, Macaque monkey, Rhesus monkey, Common marmoset, Orangutan
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Immunogen
Synthetic peptide corresponding to Human Histone H2B aa 100 to the C-terminus (C terminal) conjugated to keyhole limpet haemocyanin.
(Peptide available asab16101) -
Positive control
- WB: Calf thymus histone preparation, HeLa whole cell lysate, HeLa nuclear lysate and Histone H2B recombinant protein. ICC/IF: methanol fixed HepG2 cells. IHC-P: Human breast fibroadenoma tissue.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituents: PBS, 3% BSA
This product may contain up to 3% BSA depending on the batch. For specific batch formulations please contact us. -
Concentration information loading... -
Purity
Immunogen affinity purified -
Clonality
Polyclonal -
Isotype
IgY
Associated products
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Immunizing Peptide (Blocking)
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Isotype control
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Recombinant Protein
Applications
Our Abpromise guarantee covers the use of ab134211 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| IHC-P | Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. | |
| ICC/IF | Use a concentration of 0.1 µg/ml. | |
| WB | Use a concentration of 0.5 µg/ml. Detects a band of approximately 17 kDa (predicted molecular weight: 14 kDa). Block using 3% milk. |
Target
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Relevance
Core 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. -
Cellular localization
Nuclear -
Database links
- Entrez Gene: 337874 Human
- Entrez Gene: 54145 Human
- Entrez Gene: 8349 Human
- Entrez Gene: 104021 Mouse
- Entrez Gene: 64647 Rat
- SwissProt: P62808 Cow
- SwissProt: P02283 Drosophila melanogaster
- SwissProt: O60814 Human
see all -
Alternative names
- GL105 antibody
- H2B GL105 antibody
- H2B histone family member O antibody
see all
Images
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Immunocytochemistry/ Immunofluorescence - Anti-Histone H2B antibody (ab134211)ICC/IF image of ab134211 stained HepG2 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab134211 at 0.1µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- chicken IgY (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
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Western blot - Anti-Histone H2B antibody (ab134211)All lanes : Anti-Histone H2B antibody (ab134211) at 1 µg/ml
Lane 1 : Calf Thymus Histone Preparation Nuclear Lysate at 0.2 µg
Lane 2 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg
Lane 3 : HeLa Histone Preparation Nuclear Lysate at 2.5 µg
Lane 4 : Histone H2B Recombinant Protein at 20 µg
Lane 5 : Histone H3.1 Recombinant Protein at 0.1 µg
Secondary
All lanes : Goat Anti-Chicken IgY H&L (HRP) (ab6877) at 1/3000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 14 kDa
Exposure time: 2 minutes
This blot was produced using a 10% 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 ab134211 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H2B antibody (ab134211)IHC image of Histone H2B staining in human breast fibroadenoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol B. 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 ab134211, 1µg/ml, for 15 mins at room temperature. A goat anti-chicken biotinylated secondary antibody was used to detect the primary, and visualized using an HRP conjugated ABC system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
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.
Protocols
Datasheets and documents
References
This product has been referenced in:
- Kounatidou E et al. A novel CRISPR-engineered prostate cancer cell line defines the AR-V transcriptome and identifies PARP inhibitor sensitivities. Nucleic Acids Res 47:5634-5647 (2019). Read more (PubMed: 31006810) »
- Brinkmann V et al. Immunodetection of NETs in Paraffin-Embedded Tissue. Front Immunol 7:513 (2016). Read more (PubMed: 27920776) »
