Anti-Histone H2A (acetyl K5) antibody - ChIP Grade (ab195486)

Overview

  • Product name
    Anti-Histone H2A (acetyl K5) antibody - ChIP Grade
    See all Histone H2A primary antibodies
  • Description
    Rabbit polyclonal to Histone H2A (acetyl K5) - ChIP Grade
  • Host species
    Rabbit
  • Tested applications
    Suitable for: Dot blot, ICC/IF, CHIPseq, WB, ChIPmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human Histone H2A (acetyl K5) conjugated to keyhole limpet haemocyanin. The exact sequence is proprietary.

  • Positive control
    • Histone extracts and whole cell extracts from HeLa cells; HeLa cells; chromatin from HeLa and HeLaS3 cells.

Properties

Applications

Our Abpromise guarantee covers the use of ab195486 in the following tested applications.

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

Application Abreviews Notes
Dot blot 1/5000.
ICC/IF 1/500.
CHIPseq Use at an assay dependent concentration.

Use 0.5 μg per CHIPseq reaction.

WB 1/1000. Predicted molecular weight: 14 kDa.
ChIP Use at an assay dependent concentration.

Use 0.5-5 µg per ChIP

Target

  • Function
    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.
  • Sequence similarities
    Belongs to the histone H2A family.
  • Post-translational
    modifications
    The chromatin-associated form is phosphorylated on Thr-121 during mitosis.
    Deiminated on Arg-4 in granulocytes upon calcium entry.
    Monoubiquitination of Lys-120 by RING1 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. Monoubiquitination of Lys-120 by RNF2/RING2 can also be induced by ultraviolet and may be involved in DNA repair. 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. Monoubiquitination and ionizing radiation-induced 'Lys-63'-linked ubiquitination are distinct events.
    Phosphorylation on Ser-2 is enhanced during mitosis. Phosphorylation on Ser-2 by RPS6KA5/MSK1 directly represses transcription. Acetylation of H3 inhibits Ser-2 phosphorylation by RPS6KA5/MSK1.
    Symmetric dimethylation on Arg-4 by the PRDM1/PRMT5 complex may play a crucial role in the germ-cell lineage.
  • Cellular localization
    Nucleus. Chromosome.
  • Information by UniProt
  • Database links
  • Alternative names
    • H2A antibody
    • H2A1B_HUMAN antibody
    • H2AFM antibody
    • HIST1H2A antibody
    • HIST1H2AE antibody
    • Histone H2A type 1-B/E antibody
    • Histone H2A.2 antibody
    • Histone H2A/a antibody
    • Histone H2A/m antibody
    see all

Images

  • ChIP was performed on sheared chromatin from 1.5 million HeLaS3 cells using 0.5 μg of ab195486. The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. This figure shows the enrichment along the complete sequence and a 1 Mb region of the X-chromosome (panels A and B) and in genomic regions of chromosome 7, surrounding the ACTB gene, and of chromosome 12, surrounding the GAPDH gene (panels C and D). The position of the amplicon used for ChIP-qPCR is indicated by an arrow.

  • Immunofluorescent analysis of HeLa cells (4% formaldehyde-fixed) labeling Histone H2A (acetyl K5) with ab195486 at 1/500 dilution followed by an anti-rabbit antibody conjugated to Alexa488 (left panel). The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.

  • All lanes : Anti-Histone H2A (acetyl K5) antibody - ChIP Grade (ab195486) at 1/1000 dilution

    Lane 1 : whole cell HeLa extracts at 25 µg
    Lane 2 : HeLa histone extracts at 15 µg
    Lane 3 : recombinant histone H2A at 1 µg
    Lane 4 : recombinant histone H2B at 1 µg
    Lane 5 : recombinant histone H3 at 1 µg
    Lane 6 : recombinant histone H4 at 1 µg

    Predicted band size: 14 kDa

  • Dot Blot analysis was performed to test the cross reactivity of ab195486 against Histone H2A (acetyl K5) with peptides containing other histone modifications and the unmodified H2AK5. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane.  ab195486 was used at 1/5,000 dilution. 

  • ChIP analysis using HeLa cells, ab195486 and optimized PCR primer sets for qPCR. ChIP was performed using sheared chromatin from 1.5 million cells. A titration of the antibody consisting of 0.5, 1, 2 and, 5 µg per ChIP experiment was analysed. IgG (1 µg/IP) was used as negative IP control. QPCR was performed with primers for a region approximately 1 kb upstream of the GAPDH and ACTB promoters, used as positive controls, and for the coding region of the inactive MYOD1 gene and the Sat2 satellite repeat, used as negative controls.

References

ab195486 has not yet been referenced specifically in any publications.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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