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H2Apan polyclonal antibody

カタログ番号
フォーマット
価格
C15410166
(pAb-166-050)
50 μg
$325.00
  Bulk order
ブローカー情報



Polyclonal antibody raised in rabbit against histone H2A using a KLH-conjugated synthetic peptide containing an unmodified sequence from the C-terminal part of the protein.

LotA2371-0041
Concentration4 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 * 1-2 μg/ChIP Fig 1
ELISA 1:100-1:1,000 Fig 2
Western Blotting 1:2,000 Fig 3
Immunofluorescence 1:500 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

    H2Apan Antibody ChIP Grade

    Figure 1. ChIP results obtained with the Diagenode antibody directed against H2Apan
    ChIP assays were performed using human HeLa cells, the Diagenode antibody against H2Apan (cat. No. C15410166) and optimized PCR primer sets for qPCR. ChIP was performed with the Auto Histone ChIP-seq” kit (cat. No. C01010022) on sheared chromatin from 1 million cells using the IP-Star automated system. A titration of the antibody consisting of 1, 2, 5, and 10 μg per ChIP experiment was analysed. IgG (5 μg/IP) was used as negative IP control. QPCR was performed with primers for the GAPDH and EIF4A2 promoters, used as negative controls and for the inactive MYOD1 gene and the Sat2 satellite repeat, used as 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).

    H2Apan Antibody ELISA validation

    Figure 2. Determination of the titer
    To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H2Apan (cat. No. C15410166) in antigen coated wells. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:32,500.

    H2Apan Antibody validated in Western blot

    Figure 3. Western blot analysis using the Diagenode antibody directed against H2Apan
    Western blot was performed on whole cell (25 μg, lane 1) and histone extracts (15 μg, lane 2) from HeLa cells, and on 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Diagenode antibody against H2Apan (cat. No. C15410166). The antibody was 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.

    H2Apan Antibody validated in Immunofluorescence

    Figure 4. Immunofluorescence using the Diagenode antibody directed against H2Apan
    HeLa cells were stained with the Diagenode antibody against H2Apan (Cat. C15410166) 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 labeled with the H2Apan 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. Histones play a central role in the regulation of transcription, DNA repair, DNA replication and chromosomal stability. These different functions are established via a complex set of post-translational modifications which either directly or indirectly alter chromatin structure and DNA accessibility to facilitate transcriptional activation or repression or other nuclear processes.

  •  実験手法
    ELISA
    Enzyme-linked immunosorbent assay. 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
    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
  •  資料
    Datasheet H2Apan pAb-166-050 DATASHEET
    Datasheet description
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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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    Epigenetic Antibodies Brochure BROCHURE
    More than in any other immuoprecipitation assays, quality antibodies are critical tools in many e...
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  •  出版物

    How to properly cite this product in your work

    Diagenode strongly recommends using this: H2Apan polyclonal antibody (Diagenode Cat# C15410166 Lot# A2371-0041). Click here to copy to clipboard.

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