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H4pan Antibody - ChIP Grade (sample size)

Catalog Number
Format
Price
C15410156-10
10 µg
$95.00
  Bulk order
Other format



Polyclonal antibody raised in rabbit against histone H4 using a KLH-conjugated synthetic peptide containing an unmodified sequence from the central part of the protein.

LotA1217P
Concentration1.3 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/IP Fig 1
ELISA 1:100 Fig 2
Western Blotting 1:1,000 Fig 3
Immunofluorescence 1:1,000 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

    H4pan Antibody ChIP Grade

    Figure 1 ChIP results obtained with the Diagenode antibody directed against H4
    ChIP assays were performed using human HeLa cells, the Diagenode antibody against H4 (Cat. No. C15410156) and optimized PCR primer pairs for qPCR. ChIP was performed with the “Auto Histone ChIP-seq” kit (Cat. No. C01010022), using sheared chromatin from 1 million cells on the IP-Star Compact automated system. 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 primers specific for the promoters of the active GAPDH and c-fos genes, and for the inactive MYOD1 gene and the Sat2 satellite repeat. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

    H4pan 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 H4 (Cat. No. C15410156) in antigen coated wells. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:3,000.

    H4pan Antibody validated in Western Blot

    Figure 3 Western blot analysis using the Diagenode antibody directed against H4
    Western blot was performed on whole cell extracts (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 H4 (Cat. No. C15410156). 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.

    H4pan Antibody validated in Immunofluorescence

    Figure 4. Immunofluorescence using the Diagenode antibody directed against H4
    HeLa cells were stained with the Diagenode antibody against H4 (Cat. No. C15410156) and with DAPI. Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 1% BSA. The cells were immunofluorescently labeled with the H4 antibody (middle) diluted 1:1,000 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The left 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.

  •  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
    ELISA
    Enzyme-linked immunosorbent assay. Read more
    ChIP-qPCR (ab)
    Read more
  •  Documents
    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
    Datasheet H4pan pAb-156-050 DATASHEET
    Datasheet description
    Download
  •  Publications

    How to properly cite this product in your work

    Diagenode strongly recommends using this: H4pan Antibody - ChIP Grade (sample size) (Diagenode Cat# C15410156-10 Lot# A1217P). Click here to copy to clipboard.

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