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H3.3 Antibody - ChIP-seq Grade (sample size)

Catalog Number
Format
Price
C15210011-10
10 µg
$95.00
  Bulk order
Other format



Monoclonal antibody raised in rabbit against histone variant H3.3, using a KLH-conjugated synthetic peptide.

Lot001
Concentration1 µg/µl
Species reactivityHuman. Other species not tested.
TypeMonoclonal
PurityProtein A purified monoclonal antibody in PBS containing 50% glycerol, 1% BSA and 0.09% azide.
HostRabbit
Storage ConditionsStore at -20°C
PrecautionsThis product is for research use only. Not for use in diagnostic or therapeutic procedures.
Applications Suggested dilution References
ChIP/ChIP-seq * 4 μg/ChIP Fig 1, 2
Western Blotting 1:500 Fig 3

* Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 0.5-5 μg per IP.

  • Validation data

    H3.3 Antibody ChIP Grade

    Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H3.3
    ChIP assays were performed using human HeLa cells, the Diagenode antibody against H3.3 (cat. No. C15210011) and optimized PCR primer sets for qPCR. ChIP was performed with the "iDeal ChIP-seq" kit (cat. No. C01010055) on sheared chromatin from 1,000,000 cells. A titration of the antibody consisting of 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 the coding regions of the active CCT5 and EIF2S3 genes, used as positive controls, and for the inactive MYOD1 and TSH2B genes, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

    A.
    H3.3 Antibody ChIP-seq Grade

    B.
    H3.3 Antibody for ChIP-seq

    C.
    H3.3 Antibody for ChIP-seq assay

    D.
    H3.3 Antibody validated in ChIP-seq

    Figure 2.ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H3.3
    ChIP was performed with 1 µg of the Diagenode antibody against H3.3 (cat. No. C15210011) on sheared chromatin from 1,000,000 HeLa cells using the “iDeal ChIP-seq” kit as described above. The IP'd DNA was subsequently analysed on an Illumina HiSeq 2000. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 50 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the peak distribution along the complete sequence and a 3.5 Mb region of human chromosome 1 (figure 2A and B) and in two genomic regions surrounding the CCT5 and EIF2S3 positive control genes (figure 2C and D).

    H3.3 Antibody validated in Western Blot

    Figure 3. Western blot analysis using the Diagenode monoclonal antibody directed against H3.3
    Western blot was performed on 1 µg of recombinant histone H3.1 (lane 2) and on 1 µg of recombinant histone H3.3 (lane 1) using the Diagenode monoclonal antibody against H3.3 (cat. No. C15210011) diluted 1:500 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.

  • Target Description

    Histones are the main constituents of the protein part of chromosomes of eukaryotic cells. They are rich in the amino acids arginine and lysine and have been greatly conserved during evolution. Histones pack the DNA into tight masses of chromatin. Two core histones of each class H2A, H2B, H3 and H4 assemble and are wrapped by 146 base pairs of DNA to form one octameric nucleosome. Histone tails undergo numerous post-translational modifications, which either directly or indirectly alter chromatin structure to facilitate transcriptional activation or repression or other nuclear processes. In addition to the genetic code, combinations of the different histone modifications reveal the socalled "histone code". Histone methylation and demethylation is dynamically regulated by respectively histone methyl transferases and histone demethylases. Histone variant H3.3 is preferably present at active genes.

  •  Applications
    WB
    Western blot : The quality of antibodies used in this technique is crucial for correct and specific protein identification. Diagenode offers huge selection of highly sensitive and specific western blot-validated antibodies. Learn more about: Load... Read more
    ChIP-seq (ab)
    Read more
    ChIP-qPCR (ab)
    Read more
  •  Documents
    H3.3 monoclonal antibody DATASHEET
    Monoclonal antibody raised in rabbit against histone variant H3.3, using a KLH-conjugated synthet...
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  •  Publications

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