Diagenode

H3pan polyclonal antibody - Classic (sample size)

RFX5-polyclonal-antibody-diagenode
Catalog Number
Format
Price
C15410324-10
10 µg
$80.00
  Bulk order
Other format

Polyclonal antibody raised in rabbit against histone H3, using three KLH-conjugated synthetic peptides containing unmodified sequences from the central part and the C-terminus of the protein.

LotA2565P
Concentration3.1 μg/μl
Species reactivityHuman
TypePolyclonal
PurityAffinity 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 1-2 μg/ChIP Fig 1
ELISA 1:10,000 Fig 2
Western Blotting 1:1,000 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

    ChIP

    Figure 1. ChIP results obtained with the Diagenode antibody directed against H3
    ChIP assays were performed using human HeLa cells, the Diagenodeantibody against H3 (Cat. No. C15410324) 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 (2 μg/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the promoters of the active GAPDH and EIF4A2 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). immunoprecipitated DNA compared to input DNA after qPCR analysis).

    ELISA

    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 directed against H3pan (Cat. No. C15410324). The plates were coated with the peptides used for immunization. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be >1:1,000,000.

    Western blot

    Figure 3. Western blot analysis using the Diagenode antibody directed against H3
    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 H3 (Cat. No. C15410324). 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.

    IF

    Figure 4. Immunofluorescence using the Diagenode antibody directed against H3
    HeLa cells were stained with the Diagenode antibody against H3 (Cat. No. C15410324) 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 H3 antibody (middle) diluted 1:500 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.

  • Applications
    ChIP-qPCR (ab)
    Read more
    ELISA
    Enzyme-linked immunosorbent assay. 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
  • Documents
    H3pan polyclonal antibody DATASHEET
    Datasheet of the H3pan polyclonal antibody
    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
  • Publications

    How to properly cite this product in your work

    Diagenode strongly recommends using this: H3pan polyclonal antibody - Classic (sample size) (Diagenode Cat# C15410324-10 Lot# A2565P). Click here to copy to clipboard.

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