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Immunocytochemistry - Histone H3 (tri methyl K4) antibody [mAbcam 1012] (ab1012)
![Immunocytochemistry - Histone H3 (tri methyl K4) antibody [mAbcam 1012] (ab1012)](/ps/datasheet/images/ab1012_3.jpg)
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ChIP - Histone H3 (tri methyl K4) antibody [mAbcam 1012] - ChIP Grade (ab1012)
![ChIP - Histone H3 (tri methyl K4) antibody [mAbcam 1012] - ChIP Grade (ab1012)](/ps/datasheet/images/ab1012_5a.jpg)
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Western blot - Histone H3 (tri methyl K4) antibody [mAbcam 1012] - ChIP Grade (ab1012)
![Western blot - Histone H3 (tri methyl K4) antibody [mAbcam 1012] - ChIP Grade (ab1012)](/ps/datasheet/images/1/ab1012/Histone-H3-Primary-antibodies-ab1012-10.jpg)
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Product Name
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Histone H3 (tri methyl K4) antibody [mAbcam1012] - 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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Mouse monoclonal [mAbcam1012] 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. (Peptide available as ab1342.)
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Reacts with
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Cow, Hu, Ms, Rat, Rice
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Predicted to react
(species key)
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Mamm, At, Ce, Dm, Sc, S pom, Shp, Xl, Zfsh
(Due to sequence homology)
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Specificity
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By ELISA the antibody binds to the tri methyl K4 peptide and at high antibody concentrations to di and mono methyl K4 peptides. It does not bind to unmodified, mono, di or tri methyl K9 or di or tri methyl K27 peptides.
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Tested applications
(see key)
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ChIP, ICC/IF, WB
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Abreviews
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Application notes
(see key)
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Recommended dilutions
ChIP: Use 5-10µg for 25µg of chromatin
ICC: Use at a concentration of 5 µg/ml.
IF: 1/100.
WB: Use at a concentration of 1 - 5 µg/ml. Detects a band of approximately 17 kDa (predicted molecular weight: 15 kDa). Can be blocked with Histone H3 peptide - tri methyl K4 (ab1342).
NOT SUITABLE for blocking with milk. Block in 5% BSA for 1 hour. Our labs have investigated the blocking conditions for this antibody and found that milk significantly decreases the signal and is therefore not a suitable blocking agent for this antibody (see image below).
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
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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.
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Histone H3 (tri methyl K4) antibody [mAbcam1012] - ChIP Grade (ab1012)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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Mouse
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Clonality
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Monoclonal
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Clone number
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mAbcam1012
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Isotype
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IgG2b
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Light chain type
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kappa
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Purity
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Protein G purified
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Storage buffer
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Preservative: 0.02% Sodium Azide
Constituents: PBS 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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1.000 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 [mAbcam1012] - 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 [mAbcam1012] - ChIP Grade images, references, products related to ab1012 and other tools.
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Histone H3 (tri methyl K4) antibody [mAbcam1012] - ChIP Grade images:
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 Immunocytochemistry - Histone H3 (tri methyl K4) antibody [mAbcam 1012] (ab1012)
SK-N-SK cells were fixed in 4% paraformaldehdye, permeabilized in 0.5% Triton X-100 and incubated with ab1012 (1/100). The antibody clearly stains the nucleus (red). The cells were counterstained with DAPI (blue). 100x magnification. Darin McDonald, Cross Cancer Institute
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ChIP - Histone H3 (tri methyl K4) antibody [mAbcam 1012] - ChIP Grade (ab1012)
Chromatin was prepared from U2OS cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10min. The ChIP was performed with 25 µg of chromatin, 5 µg of ab1012 (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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Western blot - Histone H3 (tri methyl K4) antibody [mAbcam 1012] - ChIP Grade (ab1012)
Histone H3 (tri methyl K4) antibody [mAbcam1012] - ChIP Grade (ab1012) at 2.5 µg/ml + Calf Thymus Histone Preparation Nuclear Lysate (ab121) at 0.5 µg
Secondary
Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size : 15 kDa
Observed band size : 17 kDa (why is the actual band size different from the predicted?)
Exposure time : 30 seconds
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Immunocytochemistry/ Immunofluorescence - Histone H3 (tri methyl K4) antibody [mAbcam1012] - ChIP Grade (ab1012)
![Immunocytochemistry/ Immunofluorescence - Histone H3 (tri methyl K4) antibody [mAbcam1012] - ChIP Grade (ab1012)](/ps/datasheet/images/1/ab1012/Histone-H3-Primary-antibodies-ab1012-11.jpg)
ICC/IF image of ab1012 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab1012, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% PFA fixed (10 min) HepG2, Hek293 and MCF7 cells at 5µg/ml, and in 100% methanol fixed (5 min) HeLa, Hek293, HepG2 and MCF7 cells at 5µg/ml.
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Western blot - Histone H3 (tri methyl K4) antibody [mAbcam1012] - ChIP Grade (ab1012)
![Western blot - Histone H3 (tri methyl K4) antibody [mAbcam1012] - ChIP Grade (ab1012)](/ps/datasheet/images/1/ab1012/Histone-H3-Primary-antibodies-ab1012-19.jpg)
Lane 1 : Histone H3 (tri methyl K4) antibody [mAbcam1012] - ChIP Grade (ab1012) at 1 µg/ml (Blocked in 5% BSA)
Lane 2 : Histone H3 (tri methyl K4) antibody [mAbcam1012] - ChIP Grade (ab1012) at 1 µg/ml (Blocked in 5% MILK)
Lane 1 : Calf Thymus Histone Preparation Nuclear Lysate
Lane 2 : Calf Thymus Histone Preparation Nuclear Lysate
Lysates/proteins at 0.5 µg per lane.
Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size : 15 kDa
Observed band size : 17 kDa (why is the actual band size different from the predicted?)
Exposure time : 10 minutes
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- Histone H3 (tri methyl K4) antibody [mAbcam 1012] - ChIP Grade (ab1012)
![- Histone H3 (tri methyl K4) antibody [mAbcam 1012] - ChIP Grade (ab1012)](/ps/datasheet/images/ab1012-ELISA.jpg)
ELISA using ab1012 at varying antibody concentrations.
The orange line indicates binding to the tri methyl K4 peptide ab1342.
At the dilutions shown the antibody binds weakly to the di methyl K4 peptide (ab7768 - purple) and the mono methyl K4 peptide (ab1340 - grey).
Binding to the following peptides was not seen:
unmodified aa1-13 peptide (ab2903 - brown)
di methyl K9 peptide (ab1772 - red)
mono methyl K9 peptide (ab1771 - green)
tri methyl K27 peptide (1782 - purple triangle)
di methyl K27 peptide (1781 - black circle)
tri methyl K9 peptide (1773 - blue square)
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References for Histone H3 (tri methyl K4) antibody [mAbcam1012] - ChIP Grade (ab1012)
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This product has been used in: (two most recent references)
Venkatarama T et al. Repression of zygotic gene expression in the Xenopus germline. Development 137:651-60 (2010). IHC-Fr; Xenopus laevis. PubMed: 20110330
Philipot O et al. The Core Binding Factor CBF Negatively Regulates Skeletal Muscle Terminal Differentiation. PLoS One 5:e9425 (2010). ChIP; Mouse. PubMed: 20195544
See all 4 publication references for this product.
If you publish research using ab1012 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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Histone H3 (tri methyl K4) antibody [mAbcam1012] - ChIP Grade - more information
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Price and availability of products related to Histone H3 (tri methyl K4) antibody [mAbcam1012] - ChIP Grade |
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Histone H3 (tri methyl K4) antibody [mAbcam1012] - ChIP Grade - related products:
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Histone H3 (tri methyl K4) antibody [mAbcam1012] - ChIP Grade - other tools:
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Contact Abcam with a Technical Enquiry about Histone H3 (tri methyl K4) antibody [mAbcam1012] - ChIP Grade |
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All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
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![Immunocytochemistry - Histone H3 (tri methyl K4) antibody [mAbcam 1012] (ab1012)](/index.html?datasheet=1012#ab1012_3.jpg)
![ChIP - Histone H3 (tri methyl K4) antibody [mAbcam 1012] - ChIP Grade (ab1012)](/index.html?datasheet=1012#ab1012_5a.jpg)
![Western blot - Histone H3 (tri methyl K4) antibody [mAbcam 1012] - ChIP Grade (ab1012)](/index.html?datasheet=1012#Histone-H3-Primary-antibodies-ab1012-10.jpg)
