重组Anti-Histone H4 (acetyl K16)抗体[EPR1004] (ab109463)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR1004] to Histone H4 (acetyl K16)
- Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF, ChIC/CUT&RUN-seq
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
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产品名称
Anti-Histone H4 (acetyl K16)抗体[EPR1004]
参阅全部 Histone H4 一抗 -
描述
兔单克隆抗体[EPR1004] to Histone H4 (acetyl K16) -
宿主
Rabbit -
特异性
This antibody only detects Histone H4 acetylated on Lysine 16.
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经测试应用
适用于: Flow Cyt (Intra), WB, IHC-P, ICC/IF, ChIC/CUT&RUN-seqmore details
不适用于: IP -
种属反应性
与反应: Mouse, Rat, Human -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: HeLa, C6 and mouse spleen cell lysates - treated with TSA. IHC-P: Human testis, transitional cell carcinoma and colon tissues. ICC/IF: HeLa cells treated with TSA. Flow Cyt (intra): HeLa cells. ChIC/CUT&RUN-Seq: HeLa 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. Stable for 12 months at -20°C. -
存储溶液
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 59% PBS, 0.05% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR1004 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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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ab109463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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Flow Cyt (Intra) |
1/100 - 1/200.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB | (4) |
1/1000 - 1/2000. Detects a band of approximately 11 kDa (predicted molecular weight: 11 kDa).
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IHC-P | (2) |
1/100 - 1/200. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ICC/IF | (1) |
1/100 - 1/200.
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ChIC/CUT&RUN-seq |
Use at an assay dependent concentration.
2 µg |
说明 |
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Flow Cyt (Intra)
1/100 - 1/200. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
WB
1/1000 - 1/2000. Detects a band of approximately 11 kDa (predicted molecular weight: 11 kDa). |
IHC-P
1/100 - 1/200. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ICC/IF
1/100 - 1/200. |
ChIC/CUT&RUN-seq
Use at an assay dependent concentration. 2 µg |
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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ChIC/CUT&RUN was performed using a pAG-MNAse at a final concentration of 700 ng/mL, 2.5 x 10^5 HeLa (Human cervix adenocarcinoma epithelial cell line) cells and 2 µg of ab109463 [EPR1004]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown.
Additional screenshots of mapped reads can be downloaded here.
The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.
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Lanes 1-2 : Anti-Histone H4 (acetyl K16) antibody [EPR1004] (ab109463) at 1/6000 dilution
Lanes 3-4 : Anti-Histone H4 (acetyl K16) antibody [EPR1004] (ab109463) at 1/24000 dilution
Lanes 1 & 3 : Untreated C6 (Rat glial tumor glial cell) whole cell lysate
Lanes 2 & 4 : C6 (Rat glial tumor glial cell) treated with Trichostatin A (final concentration is 500ng/ml) for 4 hours whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 11 kDa
Exposure time: 3 minutesBlocking/Diluting buffer and concentration 5% NFDM/TBST
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Immunocytochemistry/ Immunofluorescence analysis of untreated HeLa cells (top row) and HeLa+ TSA(500ng/ml, 4h) cells (middle row) labeling Histone H4 (acetyl K16) with ab109463 at 1/500. Goat anti rabbit IgG(Alexa Fluor® 488); ab150077 at 1/1000 dilution was used as the secondary antibody. Cells were fixed with 4% paraformaldehyde and permeabilised with 0.1% tritonX-100. DAPI (blue) was used as a nuclear counterstain.
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All lanes : Anti-Histone H4 (acetyl K16) antibody [EPR1004] (ab109463) at 1/1000 dilution (unpurified)
Lane 1 : HeLa cell lysates, untreated
Lane 2 : HeLa cell lysates treated with TSA
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP-labelled goat anti-rabbit at 1/2000 dilution
Predicted band size: 11 kDa
Observed band size: 11 kDa -
Anti-Histone H4 (acetyl K16) antibody [EPR1004] (ab109463) at 1/1000 dilution (unpurified) + HeLa cell lysate - treated with TSA at 10 µg
Secondary
Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 11 kDa
Observed band size: 11 kDaBlocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
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Anti-Histone H4 (acetyl K16) antibody [EPR1004] (ab109463) at 1/1500 dilution (purified) + HeLa cell lysate - treated with TSA at 10 µg
Secondary
Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 11 kDa
Observed band size: 11 kDaBlocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
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Anti-Histone H4 (acetyl K16) antibody [EPR1004] (ab109463) at 1/1000 dilution (unpurified) + C6 cell lysate - treated with TSA at 10 µg
Secondary
Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 11 kDa
Observed band size: 11 kDaBlocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
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Anti-Histone H4 (acetyl K16) antibody [EPR1004] (ab109463) at 1/1500 dilution (unpurified) + C6 cell lysate - treated with TSA at 10 µg
Secondary
Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 11 kDa
Observed band size: 11 kDaBlocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
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Anti-Histone H4 (acetyl K16) antibody [EPR1004] (ab109463) at 1/1000 dilution (purified) + Mouse spleen tissue lysate at 10 µg
Secondary
Peroxidase-conjugated goat anti-rabbit IgG (H+L)
Predicted band size: 11 kDa
Observed band size: 11 kDaBlocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
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Anti-Histone H4 (acetyl K16) antibody [EPR1004] (ab109463) at 1/1500 dilution (purified) + Mouse spleen tissue lysate at 10 µg
Secondary
Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 11 kDa
Observed band size: 11 kDaBlocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon tissue labelling Histone H4 (acetyl K16) with unpurified ab109463 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon tissue labelling Histone H4 (acetyl K16) with purified ab109463 at 1/150. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human testis tissue labelling Histone H4 with unpurified ab109463 at 1/100.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human transitional cell carcinoma labelling Histone H4 (acetly K16) with unpurified ab109463 at 1/100.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling Histone H4 (acetyl K16) with unpurified ab109463 (red) at 1/100. Cells were fixed with 4% paraformaldehyde. An Alexa Fluor® 555-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.
Control: primary antibody (1/100) and secondary antibody ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).
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Immunocytochemistry/Immunofluorescence analysis of HeLa cells treated with TSA labelling Histone H4 (acetyl K16) with unpurified ab109463 (red) at 1/100. Cells were fixed with 4% paraformaldehyde. An Alexa Fluor® 555-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.
Control: primary antibody (1/100) and secondary antibody ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).
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Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling Histone H4 (acetyl K16) with purified ab109463 (red) at 1/150. Cells were fixed with 4% paraformaldehyde. An Alexa Fluor® 555-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.
Control: primary antibody (1/150) and secondary antibody ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).
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Immunocytochemistry/Immunofluorescence analysis of HeLa cells treated with TSA labelling Histone H4 (acetyl K16) with purified ab109463 (red) at 1/150. Cells were fixed with 4% paraformaldehyde. An Alexa Fluor® 555-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.
Control: primary antibody (1/150) and secondary antibody ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).
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Overlay histogram showing HeLa cells stained with unpurified ab109463 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab109463, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluorr® 488 goat anti-rabbit IgG (H+L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
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Intracellular Flow Cytometry analysis of HeLa cells labelling Histone H4 (acetyl K16)with unpurified ab109463 (red) at 1/130. Cells were fixed with 80% methanol. A FITC-conjugated goat anti-rabbit IgG was used as the secondary antibody (1/150). A rabbit monoclonal IgG was used as the isotype control (green).
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Intracellular Flow Cytometry analysis of HeLa cells labelling Histone H4 (acetyl K16) with purified ab109463 (red) at 1/200. Cells were fixed with 80% methanol. A FITC-conjugated goat anti-rabbit IgG was used as the secondary antibody (1/150). A rabbit monoclonal IgG was used as the isotype control (green).
实验方案
数据表及文件
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SDS download
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Datasheet download
文献 (73)
ab109463 被引用在 73 文献中.
- Xue X et al. Tumour cells are sensitised to ferroptosis via RB1CC1-mediated transcriptional reprogramming. Clin Transl Med 12:e747 (2022). PubMed: 35220675
- Chakraborty S et al. Heat-induced SIRT1-mediated H4K16ac deacetylation impairs resection and SMARCAD1 recruitment to double strand breaks. iScience 25:104142 (2022). PubMed: 35434547
- Chomiak AA et al. Nde1 is required for heterochromatin compaction and stability in neocortical neurons. iScience 25:104354 (2022). PubMed: 35601919
- Mattoo AR & Jessup JM MCL-1 interacts with MOF and BID to regulate H4K16 acetylation and homologous recombination repair. Cell Biol Int 46:1196-1203 (2022). PubMed: 35661479
- Li Z et al. H3K36me2 methyltransferase NSD2 orchestrates epigenetic reprogramming during spermatogenesis. Nucleic Acids Res 50:6786-6800 (2022). PubMed: 35736136