Anti-Histone H4 (tri methyl K20) antibody - ChIP Grade (ab9053)
Key features and details
- Rabbit polyclonal to Histone H4 (tri methyl K20) - ChIP Grade
- Suitable for: IHC-P, WB, ChIP
- Reacts with: Cow, Human, Recombinant fragment
- Isotype: IgG
Overview
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Product name
Anti-Histone H4 (tri methyl K20) antibody - ChIP Grade
See all Histone H4 primary antibodies -
Description
Rabbit polyclonal to Histone H4 (tri methyl K20) - ChIP Grade -
Host species
Rabbit -
Specificity
Histone H4 tri-methylated at Lys 20. -
Tested Applications & Species
See all applications and species dataApplication Species ChIP HumanIHC-P HumanWB HumanRecombinant fragment
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Immunogen
Synthetic peptide within Human Histone H4 aa 1-100 (tri methyl K20) conjugated to keyhole limpet haemocyanin. The exact sequence is proprietary.
(Peptide available asab17567) -
General notes
Mouse: Prominent staining of a subset of centromeric or pericentromeric heterochromatin. Interphase: The cells within the culture show a considerable variability in the intensity of staining with the antibody. The relationship between trimethylation levels and cell cycle have not yet been determined but may be a contributor to the amount of methylation detected in each cell. Heterochromatic regions of the interphase nucleus are the primary sites of trimethylation observed by indirect immunofluroescence. Mitosis: Discrete chromosomal regions are labelled intensely, with lower level fluorescence throughout the remainder of the chromosome arms. Immunofluorescence staining was performed as part of the nuclear antibody characterisation program at www.cellnucleus.com. Trimenthylation at Lys20 of Histone H4 isa common hallmark of human cancer (Fraga et al. 2005).
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Images
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ChIP - Anti-Histone H4 (tri methyl K20) antibody - ChIP Grade (ab9053)
Chromatin was prepared from U2OS cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 min. The ChIP was performed with 25 µg of chromatin, 6 µl of ab9053 (blue), and 20 µl of protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Taqman approach). Primers and probes are located in the first kb of the transcribed region.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H4 (tri methyl K20) antibody - ChIP Grade (ab9053) This image is courtesy of an anonymous Abreview.Immunohistochemical staining of paraffin embedded, PFA fixed, mouse liver using ab9053 at 1/100. Heat mediated antigen retrieval was performed using sodium citrate buffer, pH 6, and the sample was permeabilized with 0.05% Triton X 100. The sample was blocked in 5% BSA for 1 hour at room temperature and was then incubated with primary antibody for 3 hours in PBS/0.05% Triton X 100 at room temperature. An Alexa Fluor® 555 donkey anyti-rabbit was used as the secondary at 1/250.
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Western blot - Anti-Histone H4 (tri methyl K20) antibody - ChIP Grade (ab9053)ab9053 is specific for Histone H4 (tri-methyl K20). This is illustrated in lane 5 where the activity of ab9053 is specifically blocked by the addition of the immunizing peptide (ab17567).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H4 (tri methyl K20) antibody - ChIP Grade (ab9053)IHC image of Histone H4 (tri methyl K20) staining in Human normal skin formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab9053, 0.2µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
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Western blot - Anti-Histone H4 (tri methyl K20) antibody - ChIP Grade (ab9053)Left panel: a fission yeast whole cell extract and recombinant (r) fission yeast histone H4.
Right panel: fission yeast whole cell extracts from strains with only a single wt (h4) or mutant (h4K20R) gene were immunoblotted.
Review by Steven Sanders submitted 13 August 2004
