重组Anti-Histone H2B (acetyl K5)抗体[EP857Y] - ChIP Grade (ab40886)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP857Y] to Histone H2B (acetyl K5) - ChIP Grade
- Suitable for: WB, ChIP-sequencing, ChIP, IHC-P, ICC/IF
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
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产品名称
Anti-Histone H2B (acetyl K5)抗体[EP857Y] - ChIP Grade
参阅全部 Histone H2B 一抗 -
描述
兔单克隆抗体[EP857Y] to Histone H2B (acetyl K5) - ChIP Grade -
宿主
Rabbit -
特异性
There is cross-reactivity with H3K27Ac (Histone H3 acetylated on Lys 27).
The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.
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经测试应用
适用于: WB, ChIP-sequencing, ChIP, IHC-P, ICC/IFmore details -
种属反应性
与反应: Mouse, Rat, Human
预测可用于: Caenorhabditis elegans, Drosophila melanogaster -
免疫原
Synthetic peptide within Human Histone H2B aa 1-100 (acetyl K5). The exact sequence is proprietary.
(Peptide available asab203469) -
阳性对照
- WB: HeLa (500ng/ml trichostatin A for 4 hours), NIH/3T3 (500ng/ml trichostatin A for 4 hours) and C6 (500ng/ml trichostatin A for 4 hours) cell lysates. IHC-P: Human hepatocellular carcinoma and urinary bladder carcinoma. ICC/IF: HeLa and A431 cells. ChIP: HeLa cells.
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常规说明
We do not guarantee IHC-P for mouse and rat.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
性能
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形式
Liquid -
存放说明
Shipped 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. -
存储溶液
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EP857Y -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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ChIP Related Products
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Isotype control
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Positive Controls
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Recombinant Protein
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab40886于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。
应用 | Ab评论 | 说明 |
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WB |
1/10000. Detects a band of approximately 14 kDa (predicted molecular weight: 14 kDa).
For unpurified use at 1/50000. |
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ChIP-sequencing |
Use 4µg for 107 cells.
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ChIP | (1) |
Use 2 µg for 25 µg of chromatin.
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IHC-P |
1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
See IHC antigen retrieval protocols. The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat. |
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ICC/IF |
1/1000.
For unpurified use at 1/250 - 1/500. |
说明 |
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WB
1/10000. Detects a band of approximately 14 kDa (predicted molecular weight: 14 kDa). For unpurified use at 1/50000. |
ChIP-sequencing
Use 4µg for 107 cells. |
ChIP
Use 2 µg for 25 µg of chromatin. |
IHC-P
1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. See IHC antigen retrieval protocols. The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat. |
ICC/IF
1/1000. For unpurified use at 1/250 - 1/500. |
靶标
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相关性
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. -
细胞定位
Nuclear -
数据库链接
- Entrez Gene: 337874 Human
- Entrez Gene: 54145 Human
- Entrez Gene: 8349 Human
- Entrez Gene: 104021 Mouse
- Entrez Gene: 64647 Rat
- SwissProt: P04255 Caenorhabditis elegans
- SwissProt: P02283 Drosophila melanogaster
- SwissProt: O60814 Human
see all -
别名
- GL105 antibody
- H2B GL105 antibody
- H2B histone family member O antibody
see all
图片
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Chromatin was prepared from HeLa cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 107 HeLa cells and 4 µg of ab40886 [EP857Y]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads.
Additional screenshots of mapped reads can be downloaded here.
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All lanes : Anti-Histone H2B (acetyl K5) antibody [EP857Y] - ChIP Grade (ab40886) at 1/10000 dilution (Purified)
Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) treated with 500ng/ml trichostatin A for 4 hours whole cell lysates
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 14 kDa
Observed band size: 14 kDaBlocking/Diluting Buffer and concentration: 5% NFDM/TBST
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Chromatin was prepared from HeLa (Human cervix adenocarcinoma epithelial cell) cells according to the Abcam X-ChIP protocol*. Cells were fixed with formaldehyde for 10 minutes.
The ChIP was performed with 25 µg of chromatin, 2µg of ab40886 (red), or 2 µg of rabbit normal IgG (gray) and 20 µl of Protein A/G sepharose beads. The immunoprecipitated DNA was quantified by real time PCR (Taqman approach for active and inactive loci, Sybr green approach for heterochromatic loci).
Primers and probes are located in the first kb of the transcribed region.
*http://www.abcam.com/resources?keywords=X%20ChIP%20protocol -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H2B (acetyl K5) antibody [EP857Y] - ChIP Grade (ab40886)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human hepatocellular carcinoma tissue sections labeling Histone H2B with purified ab40886 at 1/100 dilution (5.42 µg/mL). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody.
Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
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Immunocytochemistry/ Immunofluorescence - Anti-Histone H2B (acetyl K5) antibody [EP857Y] - ChIP Grade (ab40886)
Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) treated with 500 ng/mL TSA for 4 hours cells labeling Histone H2B with purified ab40886 at 1/1000 dilution (0.54 µg/mL). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/mL). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/mL) dilution. DAPI (blue) was used as nuclear counterstain.
PBS instead of the primary antibody was used as the secondary antibody only control.
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All lanes : Anti-Histone H2B (acetyl K5) antibody [EP857Y] - ChIP Grade (ab40886) at 1/10000 dilution (Purified)
Lane 1 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates
Lane 2 : NIH/3T3 (Mouse embryonic fibroblast) treated with 500ng/ml trichostatin A for 4 hours whole cell lysates
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 14 kDa
Observed band size: 14 kDaBlocking/Diluting Buffer and concentration: 5% NFDM/TBST
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All lanes : Anti-Histone H2B (acetyl K5) antibody [EP857Y] - ChIP Grade (ab40886) at 1/10000 dilution (Purified)
Lane 1 : C6 (Rat glial tumor glial cell) whole cell lysates
Lane 2 : C6 (Rat glial tumor glial cell) treated with 500ng/ml trichostatin A for 4 hours whole cell lysates
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 14 kDa
Observed band size: 14 kDaBlocking/Diluting Buffer and concentration: 5% NFDM/TBST
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Chromatin was prepared from HeLa (Human cervix adenocarcinoma epithelial cell) cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10min. The ChIP was performed with 25µg of chromatin, 6µl of unpurified ab40886 (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 for active and inactive loci, Sybr green approach for heterochromatic loci). Primers and probes are located in the first kb of the transcribed region.
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Anti-Histone H2B (acetyl K5) antibody [EP857Y] - ChIP Grade (ab40886) at 1/3000 dilution + Drosophila melanogaster lysate (Fruit fly larvae) at 20 µg
Secondary
Anti-rabbit IgG HRP at 1/6000 dilution
Predicted band size: 14 kDa
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H2B (acetyl K5) antibody [EP857Y] - ChIP Grade (ab40886)
ab40886 (unpurified) staining human bladder carcinoma for Histone H2B expression (1/250 dilution)
Heat mediated antigen retrieval was performed with citrate buffer pH 6 before commencing with IHC staining protocol.
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Immunocytochemistry/ Immunofluorescence - Anti-Histone H2B (acetyl K5) antibody [EP857Y] - ChIP Grade (ab40886)
ab40886 (unpurified) (1/250) staining A431 (Human epidermoid carcinoma cell line) cells by immunofluorescence.
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ChIP - Anti-Histone H2B (acetyl K5) antibody [EP857Y] - ChIP Grade (ab40886)This image is courtesy of an anonymous abreview.
ab40886 (unpurified) at a 1/600 dilution for ChIP analysis of mouse dorsal skin epidermis whole tissue lysate, incubated for 15 hours at 4°C with ChIP dilution buffer. Cross-linking (X-ChIP) using 1% formaldehyde for 10 minutes.Detection step: Semiquantitative PCR.Negative control: Rabbit IgG.Cells treated with active vitamin D3.
实验方案
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.
数据表及文件
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SDS download
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Datasheet download
文献 (14)
ab40886 被引用在 14 文献中.
- Zhang L et al. Nucleotide variation in histone H2BL drives crossalk of histone modification and promotes tumour cell proliferation by upregulating c-Myc. Life Sci 271:119127 (2021). PubMed: 33515561
- Xia L et al. Modulation of IL-6 Expression by KLF4-Mediated Transactivation and PCAF-Mediated Acetylation in Sublytic C5b-9-Induced Rat Glomerular Mesangial Cells. Front Immunol 12:779667 (2021). PubMed: 35046941
- Zeng XJ et al. A2A R inhibition in alleviating spatial recognition memory impairment after TBI is associated with improvement in autophagic flux in RSC. J Cell Mol Med 24:7000-7014 (2020). PubMed: 32394486
- Zhang S et al. Epigenetic regulation of REX1 expression and chromatin binding specificity by HMGNs. Nucleic Acids Res 47:4449-4461 (2019). PubMed: 30838422
- Yucel N et al. Glucose Metabolism Drives Histone Acetylation Landscape Transitions that Dictate Muscle Stem Cell Function. Cell Rep 27:3939-3955.e6 (2019). PubMed: 31242425