Anti-Histone H3 (di methyl K79)抗体- ChIP Grade
参阅全部 Histone H3 一抗
兔多克隆抗体to Histone H3 (di methyl K79) - ChIP Grade
Rabbit
ab3594 detects a 17 kDa band in single lane Western Blot. Peptide inhibition in Western Blot hasn't been processed. Modification specificity is determined by Peptide Array. ab3594 binds strongly to the Histone H3 dii methyl K79. In Peptide Array ab3594 also partially binds to mono methyl K79 and tri methyl K79 peptides.
适用于: ChIP, WB, ICC/IF, IHC-P, PepArrmore details
与反应: Mouse, Cow, Human
预测可用于: Pig, Saccharomyces cerevisiae, Caenorhabditis elegans, a wide range of other species, Mammals, Silk worm, Triticum aestivum
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
ICC/IF: HeLa and HepG2 cells
Learn about ChIP assay kits, other ChIP antibodies, protocols and more in the ChIP assay guide.
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Liquid
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
pH: 7.40
Preservative: 0.02% Sodium azide
Constituents: 98.98% PBS, 1% BSA
Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
浓度
批次浓度范围 100 µg 浓度为 0.7 - 1 mg/ml
Immunogen affinity purified
多克隆
IgG
Abpromise™承诺保证使用ab3594于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。
| 应用 | Ab评论 | 说明 |
|---|---|---|
| ChIP | (2) | Use at an assay dependent concentration. |
| WB | (6) | Use at an assay dependent concentration. Predicted molecular weight: 15 kDa. |
| ICC/IF | (3) | Use a concentration of 0.1 - 1 µg/ml. |
| IHC-P | (1) | Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
| PepArr | Use a concentration of 0.2 - 0.02 µg/ml. |
Entrez Gene: 8350 Human
Entrez Gene: 8351 Human
Entrez Gene: 8352 Human
Entrez Gene: 8353 Human
Entrez Gene: 8354 Human
Entrez Gene: 8355 Human
Entrez Gene: 8356 Human
Entrez Gene: 8357 Human
Entrez Gene: 8358 Human
Entrez Gene: 8968 Human
Entrez Gene: 319152 Mouse
Entrez Gene: 319153 Mouse
Entrez Gene: 360198 Mouse
Entrez Gene: 97908 Mouse
Omim: 602810 Human
SwissProt: P68431 Human
SwissProt: P68433 Mouse
Unigene: 132854 Human
Unigene: 247813 Human
Unigene: 247814 Human
Unigene: 248176 Human
Unigene: 443021 Human
Unigene: 484990 Human
Unigene: 532144 Human
Unigene: 533292 Human
Unigene: 546315 Human
Unigene: 586261 Human
Unigene: 591778 Human
Unigene: 221301 Mouse
Unigene: 261657 Mouse
Unigene: 377874 Mouse
Unigene: 390558 Mouse
Unigene: 397328 Mouse
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H3/A antibody
H31_HUMAN antibody
H3F3 antibody
H3FA antibody
Hist1h3a antibody
HIST1H3B antibody
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HIST1H3D antibody
HIST1H3E antibody
HIST1H3F antibody
HIST1H3G antibody
HIST1H3H antibody
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HIST1H3J antibody
HIST3H3 antibody
histone 1, H3a antibody
Histone cluster 1, H3a antibody
Histone H3 3 pseudogene antibody
Histone H3.1 antibody
Histone H3/a antibody
Histone H3/b antibody
Histone H3/c antibody
Histone H3/d antibody
Histone H3/f antibody
Histone H3/h antibody
Histone H3/i antibody
Histone H3/j antibody
Histone H3/k antibody
Histone H3/l antibody
Western blot - Anti-Histone H3 (di methyl K79) antibody - ChIP Grade (ab3594)
All lanes : Anti-Histone H3 (di methyl K79) antibody - ChIP Grade (ab3594) at 1 µg/ml
Lane 1 : CTH (Calf Thymus Histone) at 0.5 µg
Lane 2 : NIH 3T3 nuclear lysate (Triton enriched) at 10 µg
Secondary
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/50000 dilution
Predicted band size: 15 kDa
Observed band size: 17 kDawhy is the actual band size different from the predicted?
Exposure time: 1 minute
Gel type: MES
Blocking buffer: 2% BSA
ChIP - Anti-Histone H3 (di methyl K79) antibody - ChIP Grade (ab3594)
Chromatin was prepared from U-2 OS (Human bone osteosarcoma epithelial cell line) 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 ab3594 (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.
Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (di methyl K79) antibody - ChIP Grade (ab3594)Tao J. et al PLoS One. 2017 Jun 20;12(6):e0179436. doi: 10.1371/journal.pone.0179436. eCollection 2017. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/
Verification of H3K79me2 antibody specificity.
The commercial H3K79me2 primary antibody was preincubated with (+) or without (−) antigen peptide (Abcam, catalog no. ab4556, v/v = 5:1) at room temperature for 1.5 h before the incubation with IVF porcine blastocysts. H3K79me2 signals were observed in blastocysts using unabsorbed primary antibody. By contrast, H3K79me2 signals were absent in blastocysts using pre-absorbed primary antibody. H3K79me2 antibody was localized with Alexa Flour 488-conjugated secondary antibody (green). DNA was stained with propidium iodide (red). Bottom panels showed the merged images (yellow) between H3K79me2 signals (green) and DNA staining (red). Scale bars = 50μm.
Western blot - Anti-Histone H3 (di methyl K79) antibody - ChIP Grade (ab3594)This image is courtesy of an anonymous Abreview
Anti-Histone H3 (di methyl K79) antibody - ChIP Grade (ab3594) at 20 µg + HeLa cell lysate at 20 µg
Secondary
HRP-conjugated Goat anti-rabbit IgG polyclonal at 1/2000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 15 kDa
Observed band size: 15 kDa
Exposure time: 1 minute
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (di methyl K79) antibody - ChIP Grade (ab3594)
IHC image of Histone H3 (di methyl K79) staining in human breast carcinoma FFPE section, performed on a 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 ab3594, 1µg/ml, for 8 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.
Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (di methyl K79) antibody - ChIP Grade (ab3594)
ab3594 staining Histone H3 (Di Methyl K79) in HeLa cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab3594 at 0.1µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).
Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
Peptide Array - Anti-Histone H3 (di methyl K79) antibody - ChIP Grade (ab3594)
All batches of ab3594 are tested in Peptide Array against peptides to different Histone H3 modifications. Six dilutions of each peptide are printed on to the Peptide Array in triplicate and results are averaged before being plotted on to a graph. Results show strong binding to Histone H3 - di methyl K79 peptide (ab4556), indicating that this antibody specifically recognises the Histone H3 - di methyl K79 modification.
ab4556 - Histone H3 - di methyl K79
ab4555 - Histone H3 - mono methyl K79
ab4557 - Histone H3 - tri methyl K79
ab4560 - Histone H4 - di methyl K79
ab1772 - Histone H3 - di methyl K9
ab4558 - Histone H3 - unmodified
Western blot - Anti-Histone H3 (di methyl K79) antibody - ChIP Grade (ab3594)
Anti-Histone H3 (di methyl K79) antibody - ChIP Grade (ab3594) at 1 µg/ml + Calf Thymus Histone Preparation Nuclear Lysate (ab121) at 0.5 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 15 kDa
Observed band size: 17 kDawhy is the actual band size different from the predicted?
Exposure time: 30 seconds
ChIP - Anti-Histone H3 (di methyl K79) antibody - ChIP Grade (ab3594)This image is courtesy of an anonymous Abreview
ChIP analysis using ab3594 binding Histone H3 in HeLa (Human epithelial adenocarcinoma cell line) cell nuclear lysate. Cells were cross-linked for 10 minutes with formaldehyde. Samples were incubated with primary antibody (5µg/µg chromatin) for 12 hours at 4°C. Protein binding was detected using real-time PCR.
Positive control: 5'UTR of transcribed gene.
Negative Control: Intergenic region.
