重组Anti-Histone H4 (acetyl K5)抗体[EP1000Y] - ChIP Grade (ab51997)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP1000Y] to Histone H4 (acetyl K5) - ChIP Grade
- Suitable for: ChIP, ELISA, WB, IHC-P, ICC/IF, IP
- Reacts with: Mouse, Rat, Human, Recombinant fragment
Related conjugates and formulations
概述
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产品名称
Anti-Histone H4 (acetyl K5)抗体[EP1000Y] - ChIP Grade
参阅全部 Histone H4 一抗 -
描述
兔单克隆抗体[EP1000Y] to Histone H4 (acetyl K5) - ChIP Grade -
宿主
Rabbit -
特异性
This antibody cross-reacts with H4K8ac and H4unmod peptides in peptide array assay. However, the cross-reactivity is minimal in ELISA when using optimized peptide concentration (images on website). Further optimization is needed to minimize the cross-reactivity in assays similar to peptide array.
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经测试应用
适用于: ChIP, ELISA, WB, IHC-P, ICC/IF, IPmore details -
种属反应性
与反应: Mouse, Rat, Human, Recombinant fragment
预测可用于: Xenopus laevis, Rice -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- HeLa, NIH/3T3, C6 cells or human brain glioma, human cervical carcinoma, human normal colon FFPE, mouse liver and rat cerebral cortex tissue. ChIP: Chromatin was prepared from MEF cells.
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常规说明
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.
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. Avoid freeze / thaw cycle. -
存储溶液
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EP1000Y -
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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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab51997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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ChIP | (3) |
Use 5 µg for 25 µg of chromatin.
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ELISA |
Use at an assay dependent concentration.
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WB | (2) |
1/500000. Detects a band of approximately 11 kDa (predicted molecular weight: 11 kDa).
For unpurified use at 1/10000- 1/50000. |
IHC-P |
1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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ICC/IF | (2) |
1/5000.
For unpurified use at 1/250- 1/500. |
IP |
1/30.
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说明 |
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ChIP
Use 5 µg for 25 µg of chromatin. |
ELISA
Use at an assay dependent concentration. |
WB
1/500000. Detects a band of approximately 11 kDa (predicted molecular weight: 11 kDa). For unpurified use at 1/10000- 1/50000. |
IHC-P
1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
ICC/IF
1/5000. For unpurified use at 1/250- 1/500. |
IP
1/30. |
靶标
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功能
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. -
序列相似性
Belongs to the histone H4 family. -
翻译后修饰
Acetylation at Lys-6 (H4K5ac), Lys-9 (H4K8ac), Lys-13 (H4K12ac) and Lys-17 (H4K16ac) occurs in coding regions of the genome but not in heterochromatin.
Citrullination at Arg-4 (H4R3ci) by PADI4 impairs methylation.
Monomethylation and asymmetric dimethylation at Arg-4 (H4R3me1 and H4R3me2a, respectively) by PRMT1 favors acetylation at Lys-9 (H4K8ac) and Lys-13 (H4K12ac). Demethylation is performed by JMJD6. Symmetric dimethylation on Arg-4 (H4R3me2s) by the PRDM1/PRMT5 complex may play a crucial role in the germ-cell lineage.
Monomethylated, dimethylated or trimethylated at Lys-21 (H4K20me1, H4K20me2, H4K20me3). Monomethylation is performed by SET8. Trimethylation is performed by SUV420H1 and SUV420H2 and induces gene silencing.
Ubiquitinated by the CUL4-DDB-RBX1 complex in response to ultraviolet irradiation. This may weaken the interaction between histones and DNA and facilitate DNA accessibility to repair proteins. Monoubiquitinated at Lys-92 of histone H4 (H4K91ub1) in response to DNA damage. The exact role of H4K91ub1 in DNA damage response is still unclear but it may function as a licensing signal for additional histone H4 post-translational modifications such as H4 Lys-21 methylation (H4K20me).
Sumoylated, which is associated with transcriptional repression. -
细胞定位
Nucleus. Chromosome. - Information by UniProt
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数据库链接
- Entrez Gene: 121504 Human
- Entrez Gene: 554313 Human
- Entrez Gene: 8294 Human
- Entrez Gene: 8359 Human
- Entrez Gene: 8360 Human
- Entrez Gene: 8361 Human
- Entrez Gene: 8362 Human
- Entrez Gene: 8363 Human
see all -
别名
- dJ160A22.1 antibody
- dJ160A22.2 antibody
- dJ221C16.1 antibody
see all
图片
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Chromatin was prepared from MCF7 breast cancer cell lysate. 5 ug total antibody was preincubated overnight on Protein A- Dynabeads and the chromatin (2X 15 cm dishes - around 15-20 million MCF7 cells) was incubated overnight with antibody-coupled beads. Beads were washed 6 times with modified RIPA buffer (50mM HEPES pH 7.6, 1mM EDTA, 0.7% Na deoxycholate, 1% NP-40, 0.5M LiCL) and one time with TE. DNA was eluted with TE containing 1% SDS, treated with RNAseA and Proteinase K, phenol-chloroform extracted and then made libraries for Illumina sequencing.
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Chromatin was prepared from MEF (Mouse embryonic fibroblast cell line) cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 5µg of ab51997 (red), and 20 μl protein A/G sepharose beads. 2μg of rabbit normal IgG was added to the beads control (grey). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).
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All lanes : Anti-Histone H4 (acetyl K5) antibody [EP1000Y] - ChIP Grade (ab51997) at 1/500000 dilution (purified)
Lane 1 : Nuclear extract of HeLa (Human epithelial cell line from cervix adenocarcinoma) treated with 7mM Sodium Butyrate for 24 hours
Lane 2 : Untreated nuclear extract of HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate
Lane 3 : NIH/3T3 (Mouse embryonic fibroblast cell line) treated with 500ng/ml Trichostatin A for 4 hours whole cell lysates
Lane 4 : Untreated NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysates
Lane 5 : C6 (Rat glial tumor cell line) treated with 500ng/ml Trichostatin A for 4 hours whole cell lysates
Lane 6 : Untreated C6 (Rat glial tumor cell line) 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: 11 kDa
Observed band size: 11 kDaBlocking and diluting buffer: 5% NFDM/TBST.
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IHC image of unpurified ab51997 staining Histone H4 (acetyl K5) in human colon formalin fixed paraffin embedded tissue sections, performed on a Leica Bond. 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 ab51997, 1/200 dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. No primary antibody was used in the negative control (shown on the inset).
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.
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Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma)treated with 500ng/m Trichostatin A for 4 hours labeling Histone H4 (acetyl K5) with purified ab51997 at 1/5000 dilution (0.1μg/ml). Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% tritonX-100. ab195889, an Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) was used to counterstain at 1/200 (2.5 μg/ml). ab150077, a Goat anti rabbit IgG(Alexa Fluor® 488) secondary antibody was used at 1/1000 dilution. PBS instead of the primary antibody was used as a control. DAPI nuclear staining.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse liver tissue sections labeling Histone H4 (acetyl K5) with purified ab51997 at 1/500 dilution (1 μg/ml). Heat mediated antigen retrieval was performed using EDTA Buffer, PH9. Hematoxylin was used to counter stain. ab97051, a Goat Anti-Rabbit IgG H&L (HRP) secondary antibody was used at 1/500 dilution. PBS instead of the primary antibody was used as the negative control.
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Direct ELISA with Histone H4 K5ac peptide, Histone H4 K8ac peptide, Histone H4 K16me1 peptide, Histone H4 K16me3 peptide, and Histone H4 unmodified peptide, all at 10ng/ml. ab51997 used as the primary antibody at a range of 0~1000ng/ml. Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) used as the secondary antibody at 1:2500 dilution.
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Direct ELISA with Histone H4 K5ac peptide, Histone H4 K8ac peptide, Histone H4 K16me1 peptide, Histone H4 K16me3 peptide, and Histone H4 unmodified peptide, all at 100ng/ml. ab51997 used as the primary antibody at a range of 0~1000ng/ml. Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) used as the secondary antibody at 1:2500 dilution.
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Direct ELISA with Histone H4 K5ac peptide, Histone H4 K8ac peptide, Histone H4 K16me1 peptide, Histone H4 K16me3 peptide, and Histone H4 unmodified peptide, all at 1000ng/ml. ab51997 used as the primary antibody at a range of 0~1000ng/ml. Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) used as the secondary antibody at 1:2500 dilution.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat cerebral cortex tissue sections labeling Histone H4 (acetyl K5) with purified ab51997 at 1/500 dilution (1 μg/ml). Heat mediated antigen retrieval was performed using EDTA Buffer, PH9. Hematoxylin was used to counter stain. ab97051, a Goat Anti-Rabbit IgG H&L (HRP) secondary antibody was used at 1/500 dilution. PBS instead of the primary antibody was used as the negative control.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cervical carcinoma tissue sections labeling Histone H4 (acetyl K5) with purified ab51997 at 1/500 dilution (1 μg/ml). Heat mediated antigen retrieval was performed using EDTA Buffer, PH9. Hematoxylin was used to counter stain. ab97051, a Goat Anti-Rabbit IgG H&L (HRP) secondary antibody was used at 1/500 dilution. PBS instead of the primary antibody was used as the negative control.
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All lanes : Anti-Histone H4 (acetyl K5) antibody [EP1000Y] - ChIP Grade (ab51997) at 1/1000000 dilution (unpurified)
Lane 1 : Untreated HeLa cells
Lane 2 : TSA treated HeLa calls
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat anti-rabbit HRP labelled (1:2000)
Predicted band size: 11 kDa
Observed band size: 11 kDa -
ab51997 (purified) at 1/30 dilution (2µg) immunoprecipitating Histone H4 (acetyl K5) in HeLa (Human epithelial cell line from cervix adenocarcinoma) treated with TSA whole cell lysate.
Lane 1 (input): HeLa (Human epithelial cell line from cervix adenocarcinoma) treated with TSA whole cell lysate 10ug
Lane 2 (+): ab51997+ HeLa (Human epithelial cell line from cervix adenocarcinoma) treated with TSA whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab51997 in HeLa (Human epithelial cell line from cervix adenocarcinoma) treated with TSA whole cell lysateFor western blotting, ab131366 VeriBlot for IP (HRP) was used for detection (1/10000).
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
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ICC/IF image of unpurtified ab51997 stained HeLa 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 (ab51997, 1/1000 dilution) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (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.
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 (108)
ab51997 被引用在 108 文献中.
- Sun B et al. Irisin reduces bone fracture by facilitating osteogenesis and antagonizing TGF-β/Smad signaling in a growing mouse model of osteogenesis imperfecta. J Orthop Translat 38:175-189 (2023). PubMed: 36439629
- Stewart-Morgan KR et al. Quantifying propagation of DNA methylation and hydroxymethylation with iDEMS. Nat Cell Biol 25:183-193 (2023). PubMed: 36635504
- Liu K et al. Histone deacetylase OsHDA706 increases salt tolerance via H4K5/K8 deacetylation of OsPP2C49 in rice. J Integr Plant Biol 65:1394-1407 (2023). PubMed: 36807738
- Min J et al. Preoperative environment enrichment preserved neuroligin 1 expression possibly via epigenetic regulation to reduce postoperative cognitive dysfunction in mice. CNS Neurosci Ther 28:619-629 (2022). PubMed: 34882968
- Van HT et al. Methyl-lysine readers PHF20 and PHF20L1 define two distinct gene expression-regulating NSL complexes. J Biol Chem 298:101588 (2022). PubMed: 35033534