Diagenode

H3K4me1 Antibody - ChIP-seq Grade (sample size)

目录号
格式
价格
C15410037-10
10 µg
$115.00
  Bulk order
其他格式



Polyclonal antibody raised in rabbit against histone H3 containing the monomethylated lysine 4 (H3K4me1), using a KLH-conjugated synthetic peptide.

LotA1657D
Concentration2.9 µg/µl
Species reactivityHuman, mouse, pig
TypePolyclonal
PurityAffinity purified polyclonal antibody.
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 * 1-2 μg/IP Fig 1, 2
ELISA 1:500 Fig 3
Dot Blotting 1:10,000 Fig 4
Western Blotting 1:500 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 1-5 μg per IP.

  • Validation Data

    A. H3K4me1 Antibody ChIP Grade

    B. H3K4me1 Antibody for ChIP

    Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K4me1
    ChIP was performed with the Diagenode antibody against H3K4me1 (cat. No. C15410037) on sheared chromatin from 500,000 HeLaS3 cells using the “iDeal ChIP-seq” kit (cat. No. C01010051). The chromatin was spiked with a panel of in vitro assembled nucleosomes, each containing a specific lysine methylation (SNAP-ChIP K-MetStat Panel, Epicypher). A titration of the antibody consisting of 0.5, 1, 2 and 5 µg per ChIP experiment was analysed. IgG (2 µg/IP) was used as negative IP control. Figure 1A. Quantitative PCR was performed with primers for a region surrounding the ACTB and GAS2L1 genes, used as positive controls, and for the promoters of the GAPDH and EIF4A2 genes, used as negative controls. The graph shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). Figure 1B. Recovery of the nucleosomes carrying the H3K4me1, H3K4me2, H3K4me3, H3K9me1, H3K27me1, H3K36me1, H4K20me1 and the unmodified H3K4 as determined by qPCR. The figure clearly shows the antibody is very specific in ChIP for the H3K4me1 modification.

    A. H3K4me1 Antibody ChIP-seq Grade

    B. H3K4me1 Antibody for ChIP-seq

    C. H3K4me1 Antibody validated in ChIP-seq

    D. H3K4me1 Antibody for ChIP-seq assay

    Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H3K4me1
    ChIP was performed as described above with 1 µg of the Diagenode antibody against H3K4me1 (Cat. No. C15410037). The IP’d DNA was subsequently analysed on 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 2A and B show the enrichment in chromosomal regions surrounding the ACTB and GAS2L1 positive control genes. The position of the amplicon used in the qPCR validation is indicated by an arrow. Figure 2C and D show the H3K4me1 signal in two 1 Mb regions of chromosome 5 and X, respectively.

    H3K4me1 Antibody ELISA Validation

    Figure 3. Determination of the titer
    To determine the titer, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3K4me1 (Cat. No. C15410037) in antigen coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 3), the titer of the antibody was estimated to be 1:20,100.

    H3K4me1 Antibody Dot Blot Validation

    Figure 4. Cross reactivity tests using the Diagenode antibody directed against H3K4me1
    A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K4me1 (Cat. No. C15410037) with peptides containing other modifications or unmodified sequences of histone H3 and H4. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:10,000. Figure 4 shows a high specificity of the antibody for the modification of interest.

    H3K4me1 Antibody validated in Western Blot

    Figure 5. Western blot analysis using the Diagenode antibody directed against H3K4me1
    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 H3K4me1 (Cat. No. C15410037). The antibody was diluted 1:500 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is shown on the right, the marker (in kDa) is shown on the left.

    H3K4me1 Antibody for Immunofluorescence

    Figure 6. Immunofluorescence using the Diagenode antibody directed against H3K4me1
    HeLa cells were stained with the Diagenode antibody against H3K4me1 (cat. C15410037) 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 H3K4me1 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.

  •  应用
    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-seq (ab)
    Read more
    ChIP-qPCR (ab)
    Read more
  •  文档
    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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    Datasheet H3K4me1 C15410037 DATASHEET
    Polyclonal antibody raised in rabbit against histone H3 containing the monomethylated lysine 4 (H...
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  •  Safety sheets
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  •  出版物

    How to properly cite this product in your work

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    Using our products in your publication? Let us know!

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