Histone H2B (acetyl K20) Recombinant Rabbit Monoclonal Antibody [PS01-46] - BSA and Azide free
Catalog# HA750592
Histone H2B (acetyl K20) Recombinant Rabbit Monoclonal Antibody [PS01-46] - BSA and Azide free
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WB
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IHC-P
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IF-Cell
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ChIP
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Human
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Mouse
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Rat
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unconjugated
Overview
Product Name
Histone H2B (acetyl K20) Recombinant Rabbit Monoclonal Antibody [PS01-46] - BSA and Azide free
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic peptide.
Species Reactivity
Human, Mouse, Rat
Validated Applications
WB, IHC-P, IF-Cell, ChIP
Target Molecular Weight
Predicted band size: 14 kDa
Positive Control
Hela treated with 500 ng/ml TSA for 4 hours whole cell lysate, human colon carcinoma tissue, human skin tissue, mouse skin tissue, rat skin tissue, Hela, NIH/3T3.
Conjugation
unconjugated
Clone Number
PS01-46
Product Features
Form
Liquid
Concentration
1mg/ml
Storage Instructions
Store at 2-8℃. Avoid freeze.
Storage Buffer
1*PBS (pH7.4).
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
Target
Function
Histone H2B is one of the 5 main histone proteins involved in the structure of chromatin in eukaryotic cells. Featuring a main globular domain and long N-terminal and C-terminal tails, H2B is involved with the structure of the nucleosomes. Histone H2B is a structural protein that helps organize eukaryotic DNA. It plays an important role in the biology of the nucleus where it is involved in the packaging and maintaining of chromosomes, regulation of transcription, and replication and repair of DNA. Histone H2B helps regulate chromatin structure and function through post-translational modifications and specialized histone variants. Acetylation and ubiquitination are examples of two post-translational modifications that affect the function of histone H2B in particular ways. Hyperacetylation of histone tails helps DNA-binding proteins access chromatin by weakening histone-DNA and nucleosome-nucleosome interactions. Furthermore, acetylation of a specific lysine residue binds to bromine-containing domains of certain transcription and chromatin regulatory proteins. This docking facilitates the recruitment of these proteins to the correct region of the chromosome. Ubiquitinated histone H2B is often found in regions of active transcription. Through the facilitation of chromatin remodeling, it stimulates transcriptional elongation and sets the stage for further modifications that regulate multiple elements of transcription.
Background References
1. Huang Y et al. Mechanistic and structural insights into histone H2A-H2B chaperone in chromatin regulation. Biochem J. 2020 Sep
2. Deng ZH et al. The Bre1/Rad6 machinery: writing the central histone ubiquitin mark on H2B and beyond. Chromosome Res. 2020 Dec
Sequence Similarity
Belongs to the histone H2A family.
Post-translational Modification
Deiminated on Arg-4 in granulocytes upon calcium entry.; Monoubiquitination of Lys-120 (H2AK119Ub) by RING1, TRIM37 and RNF2/RING2 complex gives a specific tag for epigenetic transcriptional repression and participates in X chromosome inactivation of female mammals. It is involved in the initiation of both imprinted and random X inactivation. Ubiquitinated H2A is enriched in inactive X chromosome chromatin. Ubiquitination of H2A functions downstream of methylation of 'Lys-27' of histone H3 (H3K27me). H2AK119Ub by RNF2/RING2 can also be induced by ultraviolet and may be involved in DNA repair. Monoubiquitination of Lys-120 (H2AK119Ub) by TRIM37 may promote transformation of cells in a number of breast cancers. Following DNA double-strand breaks (DSBs), it is ubiquitinated through 'Lys-63' linkage of ubiquitin moieties by the E2 ligase UBE2N and the E3 ligases RNF8 and RNF168, leading to the recruitment of repair proteins to sites of DNA damage. Ubiquitination at Lys-14 and Lys-16 (H2AK13Ub and H2AK15Ub, respectively) in response to DNA damage is initiated by RNF168 that mediates monoubiquitination at these 2 sites, and 'Lys-63'-linked ubiquitin are then conjugated to monoubiquitin; RNF8 is able to extend 'Lys-63'-linked ubiquitin chains in vitro. Deubiquitinated by USP51 at Lys-14 and Lys-16 (H2AK13Ub and H2AK15Ub, respectively) after damaged DNA is repaired. H2AK119Ub and ionizing radiation-induced 'Lys-63'-linked ubiquitination (H2AK13Ub and H2AK15Ub) are distinct events.; Phosphorylation on Ser-2 (H2AS1ph) is enhanced during mitosis. Phosphorylation on Ser-2 by RPS6KA5/MSK1 directly represses transcription. Acetylation of H3 inhibits Ser-2 phosphorylation by RPS6KA5/MSK1. Phosphorylation at Thr-121 (H2AT120ph) by DCAF1 is present in the regulatory region of many tumor suppresor genes and down-regulates their transcription.; Glutamine methylation at Gln-105 (H2AQ104me) by FBL is specifically dedicated to polymerase I. It is present at 35S ribosomal DNA locus and impairs binding of the FACT complex.; Symmetric dimethylation on Arg-4 by the PRDM1/PRMT5 complex may play a crucial role in the germ-cell lineage.; 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.
Subcellular Location
Nucleus, Chromosome.
UNIPROT
Synonyms
GL105 antibody
H2B GL105 antibody
H2B histone family member O antibody
H2B histone family member S antibody
H2B.1 antibody
H2B.1 B antibody
H2B.b antibody
H2B.c antibody
H2B.d antibody
H2B.e antibody
ExpandImages
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☑ Cell treatment (CT)
This data was developed using HA721285, the same antibody clone in a different buffer formulation.
Western blot analysis of Histone H2B (acetyl K20) on different lysates with Rabbit anti-Histone H2B (acetyl K20) antibody (HA721285) at 1/1,000 dilution.
Lane 1: Hela whole cell lysate (20 µg/Lane)
Lane 2: Hela treated with 500 ng/ml TSA for 4 hours whole cell lysate (20 µg/Lane)
Predicted band size: 14 kDa
Observed band size: 14 kDa
Exposure time: 2 minutes;
15% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721285) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/300,000 dilution was used for 1 hour at room temperature. -
This data was developed using HA721285, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Rabbit anti-Histone H2B (acetyl K20) antibody (HA721285) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721285) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
This data was developed using HA721285, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded human skin tissue with Rabbit anti-Histone H2B (acetyl K20) antibody (HA721285) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721285) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
This data was developed using HA721285, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded mouse skin tissue with Rabbit anti-Histone H2B (acetyl K20) antibody (HA721285) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721285) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
This data was developed using HA721285, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded rat skin tissue with Rabbit anti-Histone H2B (acetyl K20) antibody (HA721285) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721285) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
This data was developed using HA721285, the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of HeLa cells labeling Histone H2B (acetyl K20) with Rabbit anti-Histone H2B (acetyl K20) antibody (HA721285) at 1/50 dilution.
Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-Histone H2B (acetyl K20) antibody (HA721285) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. -
This data was developed using HA721285, the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of NIH/3T3 cells labeling Histone H2B (acetyl K20) with Rabbit anti-Histone H2B (acetyl K20) antibody (HA721285) at 1/50 dilution.
Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-Histone H2B (acetyl K20) antibody (HA721285) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. -
☑ Cell treatment (CT)
This data was developed using HA721285, the same antibody clone in a different buffer formulation.
Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells treated with 500ng/mL TSA for 4 hours with Histone H2B (acetyl K20) (HA721285) or Normal Rabbit IgG according to the ChIP protocol. The enriched DNA was quantified by real-time PCR using indicated primers. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
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