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H3K27me1 Antibody

Antibody ChIP icon

Catalog Number
Format
Price
C15410045-50
(pAb-045-050)
50 µg
$380.00
  Bulk order
Other format
C15410045-10
10 µg



Polyclonal antibody raised in rabbit against histone H3 containing the monomethylated lysine 27 (H3K27me1), using a KLH-conjugated synthetic peptide.

LotA932-00234P
Concentration1.93 µg/µl
Species reactivityHuman, Arabidopsis: positive. Other species: not tested.
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 * 1 µg/ChIP Fig 1
ELISA 1:500 Fig 2
Dot Blotting 1:20,000 Fig 3
Western Blotting 1:1,000 Fig 4
Immunofluorescence 1:1,000 Fig 5

* 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

    H3K27me1 Antibody ChIP Grade

    H3K27me1 Antibody ChIP Grade

    Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K27me1
    ChIP was performed with the Diagenode antibody against H3K27me1 (Cat. No. C15410045) on sheared chromatin from 500,000 K562 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 the coding sequence of the active GAPDH and ACTB genes, used as positive controls, and for the inactive MYOD1 and MYT1 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 H3K27me1, H3K27me2, H3K27me3, H3K4me1, H3K9me1, H3K36me1, H4K20me1 and the unmodified H3K4 as determined by qPCR. The figure clearly shows the antibody is specific in ChIP for the H3K27me1 modification with some slight cross reaction with H3K36me1 and H3K9me1 at higher concentrations.

    H3K27me1 Antibody validated in 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 H3K27me1 (Cat. No. C15410045). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 1), the titer of the purified antibody was estimated to be 1:32,900.

    H3K27me1 Antibody validated in Dot Blot

    Figure 3. Cross reactivity tests using the Diagenode antibody directed against H3K27me1
    A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K27me1 (Cat. No. C15410045) with peptides containing other modifications and unmodified sequences of histone H3 and H4. One hundred to 0.2 pmol of the peptide containing the respective histone modification 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.

    H3K27me1 Antibody validated in Western Blot

    Figure 4. Western blot analysis using the Diagenode antibody directed against H3K27me1
    Histone extracts (15 µg) from HeLa cells were analysed by Western blot using the Diagenode antibody against H3K27me1 (Cat. No. C15410045) 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.

    A. H3K27me1 Antibody validated in Immunofluorescence

    B. H3K27me1 Antibody validated in Immunofluorescence

    C. H3K27me1 Antibody validated in Immunofluorescence

    D. H3K27me1 Antibody validated for Immunofluorescence

    E. H3K27me1 Antibody validated for Immunofluorescence

    Figure 5. Immunofluorescence using the Diagenode antibody directed against H3K27me1
    Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H3K27me1 (Cat. No. C15410045) and with DAPI. Cells were fixed with 4% formaldehyde for 20’ and blocked with PBS/TX-100 containing 5% normal goat serum. Figure 5A: cells were immunofluorescently labeled with the H3K27me1 antibody (left) diluted 1:1,000 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa568 or with DAPI (right), which specifically labels DNA. Figure 5B, C, D and E: staining of the cells with the H3K27me1 antibody after incubation of the antibody with 2 ng/µl blocking peptide containing the unmodified and the mono-, di- and trimethylated H3K27, respectively.

  • 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.

  •  Applications
    ELISA
    Enzyme-linked immunosorbent assay. Read more
    DB
    Dot blotting 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
    ChIP-qPCR (ab)
    Read more
  •  Documents
    Datasheet H3K27me1 pAb-045-050 DATASHEET
    Datasheet description    		 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
  •  Safety sheets
    H3K27me1 antibody SDS GB en Download
    H3K27me1 antibody SDS US en Download
    H3K27me1 antibody SDS DE de Download
    H3K27me1 antibody SDS JP ja Download
    H3K27me1 antibody SDS BE nl Download
    H3K27me1 antibody SDS BE fr Download
    H3K27me1 antibody SDS FR fr Download
    H3K27me1 antibody SDS ES es Download
  •  Publications

    How to properly cite this product in your work

    Diagenode strongly recommends using this: H3K27me1 Antibody (Diagenode Cat# C15410045-50 Lot# A932-00234P). Click here to copy to clipboard.

    Using our products in your publication? Let us know!

    The Polycomb-Dependent Epigenome Controls β Cell Dysfunction, Dedifferentiation, and Diabetes.
    Lu TT, Heyne S, Dror E, Casas E, Leonhardt L, Boenke T, Yang CH, Sagar , Arrigoni L, Dalgaard K, Teperino R, Enders L, Selvaraj M, Ruf M, Raja SJ, Xie H, Boenisch U, Orkin SH, Lynn FC, Hoffman BG, Grün D, Vavouri T, Lempradl AM, Pospisilik JA
    To date, it remains largely unclear to what extent chromatin machinery contributes to the susceptibility and progression of complex diseases. Here, we combine deep epigenome mapping with single-cell transcriptomics to mine for evidence of chromatin dysregulation in type 2 diabetes. We find two chromatin-state signat...

    Applying the INTACT method to purify endosperm nuclei and to generate parental-specific epigenome profiles.
    Moreno-Romero J. et al.
    The early endosperm tissue of dicot species is very difficult to isolate by manual dissection. This protocol details how to apply the INTACT (isolation of nuclei tagged in specific cell types) system for isolating early endosperm nuclei of Arabidopsis at high purity and how to generate parental-specific epigenome pr...

    Parental epigenetic asymmetry of PRC2-mediated histone modifications in the Arabidopsis endosperm
    Moreno-Romero J et al.
    Parental genomes in the endosperm are marked by differential DNA methylation and are therefore epigenetically distinct. This epigenetic asymmetry is established in the gametes and maintained after fertilization by unknown mechanisms. In this manuscript, we have addressed the key question whether parentally inherited...

    A novel microscopy-based high-throughput screening method to identify proteins that regulate global histone modification levels.
    Baas R, Lelieveld D, van Teeffelen H, Lijnzaad P, Castelijns B, van Schaik FM, Vermeulen M, Egan DA, Timmers HT, de Graaf P
    Posttranslational modifications of histones play an important role in the regulation of gene expression and chromatin structure in eukaryotes. The balance between chromatin factors depositing (writers) and removing (erasers) histone marks regulates the steady-state levels of chromatin modifications. Here we describe...

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