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簡要描述:Anti-Histone H4 (tri methyl K20) antibody - ChIP Grade (ab9053)貨號:ab9053廠家:Abcam公司規(guī)格:100ug保存溫度:-20℃ 和 2~8℃貨期:現(xiàn)貨2~5天,期貨2-3周
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Anti-Histone H4 (tri methyl K20) antibody - ChIP Grade抗組蛋白H4產(chǎn)品信息:
Anti-Histone H4 (tri methyl K20) antibody - ChIP Grade
Rabbit polyclonal to Histone H4 (tri methyl K20) - ChIP Grade
Reacts with: Mouse, Rat, Cow, Human, Caenorhabditis elegans, Fruit fly (Drosophila melanogaster), Schizosaccharomyces pombe, Toxoplasma gondii
Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human Histone H4, tri methylated at K20.
Read Abcam's proprietary immunogen policy
(Peptide available as ab17567.)
In immunofluorescence: Human cells: The antibody stains a subset of cellular chromatin that is characteristically DAPI-rich (condensed, heterochromatic). The perinucleolar heterochromatin is particularly rich in trimethylated lysine 20 staining. 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).
Properties
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Applications
Our Abpromise guarantee covers the use of ab9053 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Notes |
---|---|
ChIP/Chip | ChIP/Chip: Use at an assay dependent concentration. PubMed: 19077033 |
IHC-P | IHC-P: Use at an assay dependent dilution. PubMed: 20007528 |
Electron Microscopy | EM: Use at an assay dependent dilution. PubMed: 20543957 |
IHC (PFA fixed) | IHC (PFA fixed): Use at an assay dependent dilution. PubMed: 17083276 |
IHC-Fr | IHC-Fr: Use at an assay dependent dilution. PubMed: 17712411 |
ICC/IF | ICC/IF: Use at an assay dependent dilution. PubMed: 19727073 |
Flow Cyt | Flow Cyt: 1/100. |
WB | WB: Use a concentration of 1 µg/ml. Detects a band of approximay 13 kDa (predicted molecular weight: 11 kDa).Can be blocked with Human Histone H4 (tri methyl K20) peptide (ab17567). |
ChIP | ChIP: Use 4-5µg for 106 cells. |
Target
Database links
Alternative names
Anti-Histone H4 (tri methyl K20) antibody - ChIP Grade images
- 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
ChIP - 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.
Western blot - Histone H4 (tri methyl K20) antibody - ChIP Grade (ab9053)
All lanes : Anti-Histone H4 (tri methyl K20) antibody - ChIP Grade (ab9053) at 1 µg/ml
Lane 1 : Calf thymus histone lysate
Lane 2 : Calf thymus histone lysate with Human Histone H4 (unmodified ) peptide (ab2622) at 1 µg/ml
Lane 3 : Calf thymus histone lysate with Human Histone H4 (mono methyl K20) peptide (ab17043) at 1 µg/ml
Lane 4 : Calf thymus histone lysate with Human Histone H4 (di methyl K20) peptide (ab14964) at 1 µg/ml
Lane 5 : Calf thymus histone lysate with Human Histone H4 (tri methyl K20) peptide (ab17567) at 1 µg/ml
Lane 6 : Calf thymus histone lysate with Human Histone H3 (tri methyl K4) peptide (ab1342) at 1 µg/ml
Lane 7 : Calf thymus histone lysate with Human Histone H3 (tri methyl K9) peptide (ab1773) at 1 µg/ml
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab6721) at 1/5000 dilution
Predicted band size : 11 kDa
Observed band size : 13 kDa (why is the actual band size different from the predicted?)
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.
Immunofluorescence - Histone H4 (tri methyl K20) antibody - ChIP Grade (ab9053)Michael Hendzel, Cross Cancer Inst, Edmonton, Canada
HeLa cells immunofluorescently stained with anti-Tri Methyl Lysine 20 of histone H4 (ab9053). HeLa cells were cultured on glass coverslips, fixed with 4% paraformaldehyde, and immunofluorescently labeled with anti-Tri K20 (green) at 1/500 and DAPI (red).
The image was collected using a 1.3 N.A. 40 X PlanApo Objective (Zeiss) using a Zeiss Axiovert 200M inverted fluorescent microscope and a SensiCam HQ (Cooke Corp.) 12-bit CCD. The images were collected using Universal Imaging MetaMorph v. 5.1 for Windows. A Physik Instruments piezo z-drive was used to collect images at 200 nm intervals throughout the depth of the cell nucleus. The digital images were processed to remove fluorescent background and colour composites were made from single focal planes using MetaMorph
Immunofluorescence - Histone H4 (tri methyl K20) antibody - ChIP Grade (ab9053)Michael Henzel, Cross Cancer Inst, Edmonton, Canada
A mouse 10T1/2 embryonic fibroblast immunofluorescently stained with anti Tri Methyl Lysine 20 of histone H4. Murine 10T1/2 embryonic fibroblasts were cultured on glass coverslips, fixed with paraformaldehyde and then immunofluorescently labeled using an anti-Tri Methyl K20 of histone H4 antibody at 1/500 (green). The nuclear chromatin was simultaneously visualized using DAPI (red).
The image was collected using a 1.4 N.A. 100 X PlanApo Objective (Zeiss) using a Zeiss Axiovert 200M inverted fluorescent microscope and a Photometrics Cascade 16-bit CCD. The images were collected using Universal Imaging MetaMorph v. 5.1 for Windows. A Physik Instruments piezo z-drive was used to collect images at 200 nm intervals throughout the depth of the cell nucleus. The digital images were processed to remove fluorescent background and colour composites were made from single focal planes using MetaMorph.
Immunofluorescence - Histone H4 (tri methyl K20) antibody - ChIP Grade (ab9053)Michael Henzel, Cross Cancer Inst, Edmonton, Canada
A field of HeLa cells stained with anti-Trimethy Lysine 20 of Histone H4.
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