Diagenode
| United States | My cart | 登录 / 注册 搜索

H3K9me3S10p polyclonal antibody

Antibody ChIP icon

目录号
格式
价格
C15310128
(CS-128-100)
100 µl
$380.00
  Bulk order
其他格式
C15310128-20
20 µl



Polyclonal antibody raised in rabbit against histone H3 containing trimethylated lysine 9 and the phosphorylated serine 10 (H3K9me3S10p), using a KLH-conjugated synthetic peptide.

LotA609-001
Concentrationnot determined
Species reactivityHuman
TypePolyclonal
PurityWhole antiserum
HostRabbit
PrecautionsThis product is for research use only. Not for use in diagnostic or therapeutic procedures.
Applications Suggested dilution References
ChIP * 5 μl/ChIP Fig 1
ELISA 1:1,000 - 1:10,000 Fig 2
Dot Blotting 1:1000 Fig 3
Western Blotting 1:500 Fig 4
Immunofluorescence 1:500 Fig 5
* Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 1-10 μg per IP.
  • Validation Data

    H3K9me3S10p Antibody ChIP Grade

    Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K9me3S10p
    ChIP assays were performed using human HeLa cells treated with colcemid, the Diagenode antibody against H3K9me3S10p (cat. No. CS-128-100) and optimized PCR primer sets for qPCR. ChIP was performed with the “LowCell# ChIP” kit (cat. No. kch-maglow-016), using sheared chromatin from 10,000 cells per IP. A titration of the antibody consisting of 1, 5, and 10 μl per ChIP experiment was analysed. Additionally, the same titration was analysed after incubation of the antibody with 5 nmol blocking peptide (cat. No. sp-128-050) for 1 hour at room temperature. IgG (5 μg/IP) was used as negative IP control. QPCR was performed with primers for the promoter of the active genes GAPDH (cat. No. pp-1001-050) and c-fos (cat. No. pp- 1004-050) and for the heterochromatin marker Sat2 (cat. No. pp-1040-050). Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

    H3K9me3S10p Antibody ELISA validation

    Figure 2. Determination of the titer
    To determine the titer, an ELISA was performed using a serial dilution of the Diagenode antibody directed against human H3K9me3S10p (cat. No. CS-128-100). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:87,000.

    H3K9me3S10p Antibody validated in Dot Blot

    Figure 3. Cross reactivity test using the Diagenode antibody directed against H3K9me3S10p
    A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K9me3S10p (cat. No. CS-128-100) with peptides containing other modifications and unmodified sequences of histone H3. 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 1:1,000. Figure 3 shows a high specificity of the antibody for the modification of interest.

    H3K9me3S10p Antibody validated in Western Blot

    Figure 4. Western blot analysis using the Diagenode antibody directed against H3K9me3S10p
    HeLa cells were treated with colcemid to block the cell cycle in metaphase and 15 μg of histone extracts of these cells were analysed by Western blot with the Diagenode antibody against H3K9me3S10p (cat. No. CS-128-100) diluted 1:500 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. The result of the Western analysis with the antibody is shown in lane 2; lane 1 shows the same analysis after incubation of the antibody with 5 nmol blocking peptide (cat. No. sp-128-050) for 1 hour at room temperature.

    H3K9me3S10p Antibody validated in Immunofluorescence

    Figure 5. Immunofluorescence using the Diagenode antibody directed against H3K9me3S10p
    HeLa cells were stained with the Diagenode antibody against H3K9me3S10p (cat. No. CS-128-100) 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 labelled with the H3K9me3S10p 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
    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
  •  文档
    Datasheet H3K9me3S10p CS-128-100 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
  •  出版物

    How to properly cite this product in your work

    Diagenode strongly recommends using this: H3K9me3S10p polyclonal antibody (Diagenode Cat# C15310128 Lot# A609-001). Click here to copy to clipboard.

    Using our products in your publication? Let us know!

    CpG signalling, H2A.Z/H3 acetylation and microRNA-mediated deferred self-attenuation orchestrate foetal NOS3 expression.
    Postberg J, Kanders M, Forcob S, Willems R, Orth V, Hensel KO, Weil PP, Wirth S, Jenke AC
    BACKGROUND: An adverse intrauterine environment leads to permanent physiological changes including vascular tone regulation, potentially influencing the risk for adult vascular diseases. We therefore aimed to monitor responsive NOS3 expression in human umbilical artery endothelial cells (HUAEC) and to study the unde...

活动

 查看所有活动

 

       Site map   |   Contact us   |   Conditions of sales   |   Conditions of purchase   |   Privacy policy