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Western blot - Histone H3 (tri methyl K4) antibody - ChIP Grade (ab8580)
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ChIP - Histone H3 (tri methyl K4) antibody - ChIP Grade (ab8580)
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Immunocytochemistry - Histone H3 (tri methyl K4) antibody - ChIP Grade (ab8580)
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Product Name
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Histone H3 (tri methyl K4) antibody - ChIP Grade
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Histone H3 (methylated) 抗体 (36件) 一覧
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Product type
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Primary antibodies
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Description
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Rabbit polyclonal to Histone H3 (tri methyl K4) - ChIP Grade
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Immunogen
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Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human Histone H3, tri methylated at K4.
(Peptide available as ab1342.)
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Reacts with
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Hu, Ms, Rat, At, Ce, Dm, Pig, Rb, Tet, Try c
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Predicted to react
(species key)
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Mamm, Cow, InMtj, Oik, Plnts, Sc
(Due to sequence homology)
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Specificity
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Specific for tri-methyl Lysine 4 of histone H3.
Note: weak reactivity to di methyl Lysine 4 of Histone H3.
There is no cross reactivity with Lysine 9.
This antibody reacts with tri methylated K4 within a sequence found in all mammals and a wide range of other species, including S. cerevisiae, S. pombe, N. crassa, Aspergillus nidulans, D. melanogaster, S. ocellaris, C. reinhardtii, C. elegans, Arabidopsis thaliana, Chicken, Xenopus, Zebrafish and Tobacco. The antibody will react with any species where the modification is present. Reactivity was confirmed using Human, Mouse, Cow, Indian Muntjac, Oikopleura and S. cerevisiae samples.
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Tested applications
(see key)
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ChIP, ChIP/Chip, Flow Cyt, ICC/IF, IHC-Fr, IHC-P, IP, WB
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Abreviews
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Application notes
(see key)
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Recommended dilutions
ChIP: Use 2µg for 25µg of chromatin
ChIP/Chip: Use at an assay dependent dilution.
Flow Cyt: Use at an assay dependent dilution.
ICC: Use at an assay dependent dilution.
ICC/IF: 1/100 - 1/5000.
IHC-P: Use at an assay dependent dilution (PMID 17634443).
IHC-Fr: 1/500.
IP: Use at an assay dependent concentration.
WB: 1/2000 - 1/5000. Detects a band of approximately 17 kDa (predicted molecular weight: 15 kDa). Can be blocked with Histone H3 peptide - tri methyl K4 (ab1342).
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
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Positive control
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calf thymus histone preparation
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Cellular localization
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Nuclear
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Research areas
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Chromatin and Nuclear Signaling >> ChIP'ing antibodies >> ChIP'ing antibodies
Chromatin and Nuclear Signaling >> Histones >> H3 >> Methylated
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Relevance
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Histones are basic nuclear proteins that, together with DNA, make up the nucleosome structure in eukaryotes. Nucleosomes consist of approximately 146 bp of DNA wrapped around a histone octamer, which is composed of pairs of each of the four core histones (H2A, H2B, H3, and H4). The chromatin structure is further compacted through the interaction of a linker histone H1.
Methylation of specific residues in the histone tails is a central modification for regulating epigenetic transitions in chromatin. Modified lysine residues can exist in a mono-, di-, or tri-methylated state, whilst arginine residues can be mono- or di-methylated.
In immunofluorescence, a distinct property of tri methyl lysine 4 is its apparent 'ringing' of regions that appear as nucleoplasmic 'holes'. These represent the positions of splicing factor compartments, which often are easy to identify using only DNA stains in Indian muntjac fibroblasts. These splicing factor compartments are known to be preferentially associated with active genes and highly acetylated histone H3. The antibody, as expected, fails to stain heterochromatin (work by Kirk McManus, lab of Michael Hendzel).
The immunofluorescence results suggest this antibody is an exceptional euchromatin probe.
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Histone H3 (tri methyl K4) antibody - ChIP Grade (ab8580)の別名 |
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Database links
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The links below go to external sites and will open in a new browser window |
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Raised in
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Rabbit
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Clonality
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Polyclonal
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Isotype
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IgG
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Purity
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Immunogen affinity purified
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Storage buffer
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Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4 Material safety datasheet (MSDS) for this product:
Sodium Azide MSDS
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Form
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Liquid
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Concentration
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0.300 mg/ml
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Storage instructions
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Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
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Abcam では全ての製品情報を一元管理しており、 Histone H3 (tri methyl K4) antibody - ChIP Grade について私たちが知りうる全ての情報がこのデータシートでご覧いただけます。ご不明な点がありましたら、 お問い合わせください。 |
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See below for Histone H3 (tri methyl K4) antibody - ChIP Grade images, references, products related to ab8580 and other tools.
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Histone H3 (tri methyl K4) antibody - ChIP Grade images:
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 ChIP - Histone H3 (tri methyl K4) antibody - ChIP Grade (ab8580)
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, 2 µg of ab8580 (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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Immunocytochemistry - Histone H3 (tri methyl K4) antibody - ChIP Grade (ab8580)
ab8580 staining cultured human primary fibroblasts by ICC. Cells were PFA fixed and permeabilized in TritonX100 and saponin prior to blocking with 1% BSA for 1 hour at RT. The primary antibody was diluted 1/100 and incubated with the sample for 16 hours at 4°C. A FITC-conjugated rabbit anti-rabbit IgG antibody was used as the secondary. This image is courtesy of an Abreview submitted by Dr Eva Bartova
See Abreview
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Immunofluorescence - Histone H3 (tri methyl K4) antibody - ChIP Grade (ab8580)
Staining (green) with the anti-trimethyl Lysine K4 of Histone H3 antibody (ab8580) shows ringing of regions that appear as nucleoplasmic holes. These represent the positions of splicing factor compartments, which are preferentially associated with active genes and highly acetylated histone H3.
The antibody, as expected, fails to stain heterochromatin (red). Kirk McManus in the lab of Michael Hendzel, University of Alberta
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Western blot - Histone H3 (tri methyl K4) antibody - ChIP Grade (ab8580)
All lanes : Histone H3 (tri methyl K4) antibody - ChIP Grade (ab8580) at 1/5000 dilution
Lane 1 : S. cerevisiae whole cell lysate with WT
Lane 2 : S. cerevisiae whole cell lysate with set1
Lane 3 : S. cerevisiae whole cell lysate with bre2
Lane 4 : S. cerevisiae whole cell lysate with sdc1
Lane 5 : S. cerevisiae whole cell lysate with shg1
Lane 6 : S. cerevisiae whole cell lysate with spp1
Lane 7 : S. cerevisiae whole cell lysate with swd1
Lane 8 : S. cerevisiae whole cell lysate with swd3
Lysates/proteins at 40 µg per lane.
Performed under reducing conditions.
Predicted band size : 15 kDa
Observed band size : 15 kDa
Protein resolved on 15% SDS-PAGE gel. After transfer to PVDF membrane, blots were blocked in 1X PBS, 0.1% Tween-20, and 5% milk. ab1791 was diluted in 5 ml blocking buffer at 1/5000. Blots plus primary antibodies were either incubated overnight at 4C or at RT for 2 hr. Blots were washed 6X for 10 min each in PBS with 0.1% Tween-20 before addition of secondary antibodies. Secondary antibodies were diluted 1/2,000 in blocking buffer and incubated with blots for 2 hr at RT. Secondary blots were washed 4X for 10 min each in PBS with 0.1% Tween-20 and 2X for 10 min each in PBS.
John E. Mueller and J. Ruth German (Mary Bryk lab)
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Immunofluorescence - Histone H3 (tri methyl K4) antibody - ChIP Grade (ab8580)
ab8580 use in bladder cancer cell line
IF of primary cultures of bladder cancer which arise from urothelial cells to determine whether the alterations in chromatin during cancer development would be enhanced by the histone modification antibodies. Both cell types gave very weak staining with the trimethyl-K9-H3 antibody. Wolfgang Schulz
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Immunofluorescence - Histone H3 (tri methyl K4) antibody - ChIP Grade (ab8580)
Mouse zygotes stained with the Tri-Methyl K4 histone H3 antibody
(green) and DNA (blue). This modification is readily detected in the two
pronuclei of the zygote. This image was kindly supplied as part of the review submitted by Dr Maria Elena Torres Padilla (University of Cambridge, UK).
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Immunofluorescence - Histone H3 (tri methyl K4) antibody - ChIP Grade (ab8580)
Human female lymphoblast immunostained with ab8580 (1:100)(yellowish green) specific for histone H3 lysine 4 (H3-K4) trimethylation; the DNA is stained red with propidium iodide (PI).Note the inactive X chromosome (arrow) and pericentromeric heterochromatin are largely devoid of this modification. Ahmad Khalil and Daniel Driscoll, University of Florida College of Medicine.
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References for Histone H3 (tri methyl K4) antibody - ChIP Grade (ab8580)
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This product has been used in: (two most recent references)
Hajji N et al. Opposing effects of hMOF and SIRT1 on H4K16 acetylation and the sensitivity to the topoisomerase II inhibitor etoposide. Oncogene : (2010). WB; Human. PubMed: 20118981
Maass PG et al. A cis-regulatory site downregulates PTHLH in translocation t(8;12)(q13;p11.2) and leads to Brachydactyly Type E. Hum Mol Genet 19:848-60 (2010). ChIP; Human. PubMed: 20015959
See all 228 publication references for this product.
If you publish research using ab8580 please let us know so that we can cite the reference on this datasheet.
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Search PubMed (MEDLINE) for references to Histone H3
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