Diagenode

H3K14ac monoclonal antibody - Classic

Histone-Deacetylase-polyclonal-antibody-diagenode
Catalog Number
Format
Price
C15210005
100 µg /100 µl
$295.00
  Bulk order

Monoclonal antibody raised in rabbit against histone H3 acetylated at Lys14 (H3K14ac), using a KLH-conjugated synthetic peptide.

Lot001
Concentration1 μg/μl
Species reactivityHuman
TypeMonoclonal Chip grade, ChIP-seq grade
PurityProtein A purified
HostRabbit
StorageStore at -20°C; for long storage, store at -80°C. Avoid multiple freeze-thaw cycles.
PrecautionsThis product is for research use only. Not for use in diagnostic or therapeutic procedures.
Applications Suggested dilution References
ChIP * 1 μg/ChIP Fig 1, 2
Dot blot 1:10,000 Fig 3
Western Blotting 1:2,000 Fig 4
Immunofluorescence 1:500 Fig 5

* Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 1-5 μg per IP.

  • Validation Data

    ChIP

    Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H3K14ac
    ChIP assays were performed using HeLa cells, the Diagenode antibody against H3K14ac (cat. No. C15210005) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (cat. No. C01010051), using sheared chromatin from 1 million cells. A titration consisting of 0.5, 1, 2 and 5 µg of antibody per ChIP experiment was analyzed. IgG (1 µg/IP) was used as a negative IP control. Quantitative PCR was performed with optimized primers for the promoters of the EIF2S3 and GAPDH genes, used as positive controls, and for the MYOD1 gene and the Sat2 satellite repeat, used as negative control. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

    A.ChIP-seq
    B.ChIP-seq
    C.ChIP-seq
    D.ChIP-seq

    Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H3K14ac
    ChIP was performed on sheared chromatin from 1 million HeLa cells using 1 μg of the Diagenode antibody against H3K14ac (Cat. No. C15210005) as described above. 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 50 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the enrichment along the complete sequence and a 1 Mb region of human chromosome 3 (fig 2A and B), and in two genomic regions surrounding the EIF2S3 and GAPDH positive control genes (fig 2C and D).

    Dot blotting

    Figure 3. Cross reactivity tests using the Diagenode monoclonal antibody directed against H3K14ac
    A Dot Blot analysis was performed to test the cross reactivity of the Diagenode monoclonal antibody against H3K14ac (Cat. No. C15210005) with peptides containing different modifications or unmodified sequences of histone H3. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:10,000. Figure 2 shows a high specificity of the antibody for the modification of interest.

    Western blot

    Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against H3K14ac
    Histone extracts from HeLa cells treated with butyrate (lane 2) or untreated control cells (lane 1) were analysed by Western blot using the Diagenode monoclonal antibody against H3K14ac (Cat. No. C15210005) diluted 1:2,000 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.

    Immunofluorescence

    Figure 5. Immunofluorescence using the Diagenode monoclonal antibody directed against H3K14ac
    HeLa cells were stained with the Diagenode antibody against H3K14ac (Cat. No. C15210005 red) diluted 1:500. Actin filaments were stained with fluorescein phalladoin (green).

  • Applications
    IF
    Immunofluorescence: Diagenode offers huge selection of highly sensitive antibodies validated in IF. Immunofluorescence using the Diagenode monoclonal antibody directed against CRISPR/Cas9 HeLa cells transfected with a Cas9 expression vector (... Read more
    DB
    Dot blotting Read more
    ChIP-qPCR (ab)
    Read more
    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
  • Documents
    H3K14ac monoclonal antibody DATASHEET
    Monoclonal antibody raised in rabbit against histone H3 acetylated at Lys14 (H3K14ac), using a KL...
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    Epigenetic Antibodies Brochure BROCHURE
    More than in any other immuoprecipitation assays, quality antibodies are critical tools in many e...
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    Antibodies you can trust POSTER
    Epigenetic research tools have evolved over time from endpoint PCR to qPCR to the analyses of lar...
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  • Publications

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