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H2AK119ub polyclonal antibody (sample size)

カタログ番号
フォーマット
価格
C15410002-10
10 µg
$95.00
  Bulk order
ブローカー情報



Polyclonal antibody raised in rabbit against the region of histone H2A containing the ubiquitinylated lysine 119 (H2AK119ub), using a KLH-conjugated synthetic peptide.

LotA1615-001P
Concentration1.1 µg/µl
Species reactivityHuman
TypePolyclonal
PurityAffinity purified
HostRabbit
PrecautionsThis product is for research use only. Not for use in diagnostic or therapeutic procedures.
Applications Suggested dilution References
ChIP * 2 - 5 μg/IP Fig 1
ELISA 1:200 Fig 2
Dot Blotting 1:20,000 Fig 3
Western Blotting 1:200 Fig 4
* 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

    H2AK119ub Antibody ChIP Grade

    Figure 1. ChIP results obtained with the Diagenode antibody directed against H2AK119ub.
    ChIP assays were performed using human HeLa cells, the Diagenode antibody against H2AK119ub (cat. No. C15410002) and optimized PCR primer pairs for qPCR. ChIP was performed with the “Auto Histone ChIP-seq” kit (cat. No. AB-Auto02-A100), using sheared chromatin from 1 million cells on the SX-8G IP-Star automated system. A titration consisting of 1, 2, 5 and 10 μg of antibody per ChIP experiment was analyzed. IgG (2 μg/IP) was used as a negative IP control. Quantitative PCR was performed with primers for the promoters of the active GAPDH and c-fos genes, used as negative controls, and for the inactive MYT1 and TSH2B genes, used as a positive controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

    H2AK119ub Antibody ELISA validation

    Figure 2. Determination of the antibody titer.
    To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody against H2AK119ub (cat. No. C15410002). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:23,000.

    H2AK119ub Antibody validated in Dot Blot

    Figure 3. Cross reactivity tests using the Diagenode antibody directed against H2AK119ub
    To test the cross reactivity of the Diagenode antibody against H2AK119ub (cat. No. C15410002), a Dot Blot analysis was performed with peptides containing other histone ubiquitinylations and unmodified sequences. 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 3 shows a high specificity of the antibody for the modification of interest.

    H2AK119ub Antibody validated in Western Blot

    Figure 4. Western blot analysis using the Diagenode antibody directed against H2AK119ub.
    Western blot was performed on whole cell extracts from HeLa cells (25 μg, lane 1), and on 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 2, 3, 4 and 5, respectively) using the Diagenode antibody against H2AK119ub (cat. No. C15410002). The antibody was diluted 1:200 in TBS- Tween containing 5% skimmed milk. 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 so-called “histone code”. Histone methylation and demethylation is dynamically regulated by respectively histone methyl transferases and histone demethylases. Ubiquitinylation of histone H2AK119 is associated with Polycomb mediated gene silencing.

  •  実験手法
    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
    ELISA
    Enzyme-linked immunosorbent assay. Read more
    DB
    Dot blotting Read more
    ChIP-qPCR (ab)
    Read more
  •  資料
    Datasheet H2AK119ub C15410002 DATASHEET
    Datasheet description
    Download
  •  出版物

    How to properly cite this product in your work

    Diagenode strongly recommends using this: H2AK119ub polyclonal antibody (sample size) (Diagenode Cat# C15410002-10 Lot# A1615-001P). Click here to copy to clipboard.

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