Anti-Histone H4抗体- ChIP Grade (ab18253)


  • 产品名称Anti-Histone H4抗体- ChIP Grade
    参阅全部 Histone H4 一抗
  • 描述
    兔多克隆抗体to Histone H4 - ChIP Grade
  • 经测试应用适用于: ChIP, IP, ICC/IF, IHC-P, WBmore details
  • 种属反应性
    与反应: Cow, Human
  • 免疫原

    Synthetic peptide corresponding to Human Histone H4 aa 1-100 (N terminal) conjugated to Keyhole Limpet Haemocyanin (KLH).
    Database link: P62805
    (Peptide available as ab19747)

  • 阳性对照
    • Calf Thymus Histone Preparation; Hela whole cell extract IF/ICC: HeLa cell line.



Our Abpromise guarantee covers the use of ab18253 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
ChIP Use 2 µg for 25 µg of chromatin.
IP Use a concentration of 5 µg/ml.
ICC/IF Use a concentration of 1 µg/ml.
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 15 kDa (predicted molecular weight: 11 kDa).


  • 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
  • 数据库链接
  • 别名
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Anti-Histone H4 antibody - ChIP Grade 图像

  • All lanes : Anti-Histone H4 antibody - ChIP Grade (ab18253) at 1 µg/ml

    Lane 1 : Calf Thymus Histone lysate
    Lane 2 : Calf Thymus Histone lysate with Human Histone H4 peptide (ab19747) at 1 µg/ml

    Lysates/proteins at 0.5 µg per lane.

    Alexa Fluor Goat polyclonal to Rabbit IgG at 1/10000 dilution

    Predicted band size : 11 kDa
    Observed band size : 14 kDa (why is the actual band size different from the predicted?)
  • Chromatin was prepared from HeLa cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 2µg of ab18253 (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.
  • Histone H4 was immunoprecipitated using 0.5mg Hela whole cell extract, 5ug of Rabbit polyclonal to Histone H4 and 50µl of protein G magnetic beads (lane 1). The antibody was incubated with the Protein G beads for 10min under agitation. No antibody was added to the control (lane 2). Hela whole cell extract diluted in RIPA buffer was added to each sample and incubated for 10min under agitation. Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab18253. Secondary: Clean blot (HRP conjugate) at 1/1000 dilution. Band: 14kDa: Histone H4.
  • ICC/IF image of ab18253 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 (ab18253, 1µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 Goat anti-Rabbit IgG (H+L) used at a 1/250 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.

  • IHC image of Histone H4 staining in human breast carcinoma FFPE section, performed on a BondTM system using the standard protocol F. 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 ab18253, 1µg/ml, for 8 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.

Anti-Histone H4 antibody - ChIP Grade (ab18253)参考文献

ab18253 has not yet been referenced specifically in any publications.

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