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H2A.Zac Antibody - ChIP-seq Grade

目录号
格式
价格
C15410173
(pAb-173-050)
50 μg
$332.00
  Bulk order
其他格式



Polyclonal antibody raised in rabbit against histone H2A.Z acetylated at lysines 4, 7 and 11, using a KLH-conjugated synthetic peptide.

LotA1775P
Concentration1.4 µg/µl
Species reactivityHuman: positive. Other species: not tested.
TypePolyclonal, ChIP grade, ChIP-seq grade
PurityAffinity purified
HostRabbit
Storage ConditionsStore at -20°C; for long storage, store at -80°C. Avoid multiple freeze-thaw cycles.
Storage BufferPBS containing 0.05% azide and 0.05% ProClin 300
PrecautionsThis product is for research use only. Not for use in diagnostic or therapeutic procedures.
Applications Suggested dilution References
ChIP/ChIP-seq * 0.5 μg/ChIP Fig 1, 2
ELISA 1:5,000 Fig 3
Dot Blotting 1:20,000 Fig 4
Western Blotting 1:1,000 Fig 5
Immunofluorescence 1:500 Fig 6

* 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

    Figure 1. ChIP results obtained with the Diagenode antibody directed against H2A.Zac
    ChIP assays were performed using HeLa cells, the Diagenode antibody against H2A.Zac (cat. No. C15410173) and optimized primer pairs for qPCR. ChIP was performed on sheared chromatin from 100,000 K562 cells using the iDeal ChIP-seq kit. A titration of the antibody consisting of 0.2, 0.5, 1 and 2 µg per ChIP experiment was analysed. IgG (1 µg/IP) was used as negative IP control. QPCR was performed using primers specific for the promoter of the EIF4A2 gene, used as positive control target and for the coding region of the MYT1 gene, and the Sat2 satellite repeat, used as negative control targets. Figure 1 shows the recovery (the relative amount of immunoprecipitated DNA compared to input DNA).

    A.
    B.
    C.
    D.

    Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H2A.Zac
    ChIP was performed with 0.5 µg of the Diagenode antibody against H2A.Zac (cat. No. C15410173) as described above. The IP'd DNA was subsequently analysed with an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. Figure 2 shows the peak distribution along the complete sequence and a 1 Mb region of human chromosome 1 (figure 2A and B) and in two regions surrounding the GAPDH and the EIF4A2 positive control gene (figure 2C and D, respectively).

    Figure 3. Determination of the antibody titer
    To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H2A.Zac (cat. No. C15410173). The antigen used was a peptide containing the histone modifications of interest. By plotting the absorbance against the antibody dilution (Figure 3), the titer of the purified antibody was estimated to be 1:265,000.

    Figure 4. Cross reactivity test using the Diagenode antibody directed against H2A.Zac
    A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H2A.Zac (cat. No. C15410173) with peptides containing other histone acetylations and the unmodified H2A.Z sequence. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 4 shows a high specificity of the antibody for the modification of interest.

    Figure 5. Western blot analysis using the Diagenode antibody directed against H2A.Zac
    Western blot was performed on whole cell extracts (25 µg, lane 1) from HeLa cells, and on 1 µg of recombinant histone H2A, H2B, H3 and H4 (lane 2, 3, 4 and 5, respectively) using the Diagenode antibody against H2A.Zac (cat. No. C15410173). The antibody was diluted 1:1,000 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.

    Figure 6. Immunofluorescence using the Diagenode antibody directed against H2A.Zac
    HeLa cells were stained with the Diagenode antibody against H2A.Zac (cat. No. C15410173) and with DAPI. Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the H2A.Zac antibody (left) diluted 1:500 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.

  • 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 so-called "histone code". Histone methylation and demethylation is dynamically regulated by respectively histone methyl transferases and histone demethylases. Acetylation of the histone H2A variant H2A.Z is associated with the promoters of active genes.

  •  应用
    ELISA
    Enzyme-linked immunosorbent assay. Read more
    DB
    Dot blotting Read more
    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
    ChIP-seq (ab)
    Read more
    ChIP-qPCR (ab)
    Read more
  •  文档
    Datasheet H2AZac C15410173 DATASHEET
    Polyclonal antibody raised in rabbit against histone H2A.Z acetylated at lysines 4, 7 and 11, usi...
    Download
    Antibodies you can trust POSTER
    Epigenetic research tools have evolved over time from endpoint PCR to qPCR to the analyses of lar...
    Download
    Epigenetic Antibodies Brochure BROCHURE
    More than in any other immuoprecipitation assays, quality antibodies are critical tools in many e...
    Download
  •  Safety sheets
    H2A.Zac Antibody - ChIP-seq Grade SDS US en Download
    H2A.Zac Antibody - ChIP-seq Grade SDS GB en Download
    H2A.Zac Antibody - ChIP-seq Grade SDS ES es Download
    H2A.Zac Antibody - ChIP-seq Grade SDS DE de Download
    H2A.Zac Antibody - ChIP-seq Grade SDS JP ja Download
    H2A.Zac Antibody - ChIP-seq Grade SDS BE nl Download
    H2A.Zac Antibody - ChIP-seq Grade SDS BE fr Download
    H2A.Zac Antibody - ChIP-seq Grade SDS FR fr Download
  •  出版物

    How to properly cite this product in your work

    Diagenode strongly recommends using this: H2A.Zac Antibody - ChIP-seq Grade (Diagenode Cat# C15410173 Lot# A1775P). Click here to copy to clipboard.

    Using our products in your publication? Let us know!

    Neonatal exposure to hyperoxia leads to persistent disturbances in pulmonary histone signatures associated with NOS3 and STAT3 in a mouse model.
    Chao CM, van den Bruck R, Lork S, Merkle J, Krampen L, Weil PP, Aydin M, Bellusci S, Jenke AC, Postberg J
    Background: Early pulmonary oxygen exposure is one of the most important factors implicated in the development of bronchopulmonary dysplasia (BPD). Methods: Here, we analyzed short- and long-term effects of neonatal hyperoxia on NOS3 and STAT3 expression and corresponding epigenetic signatures using a hyperoxia-base...

    CpG signalling, H2A.Z/H3 acetylation and microRNA-mediated deferred self-attenuation orchestrate foetal NOS3 expression.
    Postberg J, Kanders M, Forcob S, Willems R, Orth V, Hensel KO, Weil PP, Wirth S, Jenke AC
    BACKGROUND: An adverse intrauterine environment leads to permanent physiological changes including vascular tone regulation, potentially influencing the risk for adult vascular diseases. We therefore aimed to monitor responsive NOS3 expression in human umbilical artery endothelial cells (HUAEC) and to study the unde...

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