Anti-Histone H3 (mono methyl K9)抗体- ChIP Grade
参阅全部 Histone H3 一抗
兔多克隆抗体to Histone H3 (mono methyl K9) - ChIP Grade
Rabbit
Weak cross reactivity is observed with mono methyl K27 Histone H3. No cross-reactivity is seen with di or tri methyl K27.
适用于: IP, WB, ChIP, ICC/IFmore details
与反应: Cow, Human, Arabidopsis thaliana
预测可用于: Mouse, Rat, Saccharomyces cerevisiae, Xenopus laevis, Indian muntjac, Schizosaccharomyces pombe, Mammals
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
(Peptide available as ab1771)
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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: 1% BSA, 98.98% PBS
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
100 µg 浓度为 1 mg/ml
Immunogen affinity purified
多克隆
IgG
Abpromise™承诺保证使用ab9045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 |
|---|---|---|
| IP | Use at an assay dependent concentration. | |
| WB | (7) | 1/1000. Detects a band of approximately 15 kDa (predicted molecular weight: 17 kDa). Can be blocked with Histone H3 peptide - mono methyl K9 (ab1771). |
| ChIP | (1) | Use 4-5µg for 106 cells. |
| ICC/IF | (4) | Use at an assay dependent concentration. |
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
Unigene: 138090 Rat
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ChIP - Anti-Histone H3 (mono methyl K9) antibody - ChIP Grade (ab9045)
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 ab9045 (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 for active and inactive loci, Sybr green approach for heterochromatic loci). Primers and probes are located in the first kb of the transcribed region.
Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (mono methyl K9) antibody - ChIP Grade (ab9045)This image is courtesy of Kirk McManus in the lab of Michael Hendzel, Univeristy of Alberta
Anti-mono methyl lysine 9 of histone H3 (green) has a distribution often associated with euchromatic probes (small foci). Most of these foci localize to regions that contain obvious enrichments of DNA with DAPI staining (red). The perinucleolar chromatin is typically a site enriched in monomethylated lysine 9.
Top left: Mono-methyl Lys 9 (ab9045); Bottom left: DAPI; Top right: Merge of ab9045 (green) and DAPI (red).
Western blot - Anti-Histone H3 (mono methyl K9) antibody - ChIP Grade (ab9045)
All lanes : Anti-Histone H3 (mono methyl K9) antibody - ChIP Grade (ab9045) at 1 µg/ml
Lane 2 : Human Histone H3 (unmodified) peptide (ab7228)
Lane 3 : Human Histone H3 (mono methyl K27) peptide (ab1780)
Lane 4 : Human Histone H3 (di methyl K27) peptide (ab1781)
Lane 5 : Human Histone H3 (tri methyl K27) peptide (ab1782)
Lane 6 : Human Histone H3 (mono methyl K4) peptide (ab1340)
Lane 7 : Human Histone H3 (mono methyl K9) peptide (ab1771)
Predicted band size: 17 kDa
Rabbit polyclonal to Histone H3 K9 Methyl K9 (1/1000)
Peptides at 1 ug/ml
1XTBS, 5%BSA, 0.5% Tween
This antibody shows significantly greater reactivity with mono methyl K9. This can be seen in lane 7, as the addition of ab1771 (mono methyl K9) completely blocks the activity of ab9045. Weaker cross-reactivity is seen against mono methyl K27. This is shown in lane 3, as the addition of ab1780 only partially blocks the activity of ab9045.
Western blot - Anti-Histone H3 (mono methyl K9) antibody - ChIP Grade (ab9045)
All lanes : Anti-Histone H3 (mono methyl K9) antibody - ChIP Grade (ab9045) at 1 µg/ml
Lane 1 : Calf Thymus Histone Preparation Nuclear Lysate (ab121)
Lane 2 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) with Human Histone H3 (unmodified) peptide (ab7228) at 0.5 µg/ml
Lane 3 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) with Human Histone H3 (mono methyl K4) peptide (ab1340) at 0.5 µg/ml
Lane 4 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) with Human Histone H3 (di methyl K4) peptide (ab7768) at 0.5 µg/ml
Lane 5 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) with Human Histone H3 (tri methyl K4) peptide (ab1342) at 0.5 µg/ml
Lane 6 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) with Human Histone H3 (mono methyl K9) peptide (ab1771) at 0.5 µg/ml
Lane 7 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) with Human Histone H3 (di methyl K9) peptide (ab1772) at 0.5 µg/ml
Lane 8 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) with Human Histone H3 (tri methyl K9) peptide (ab1773) at 0.5 µg/ml
Lane 9 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) with Human Histone H3 (mono methyl K27) peptide (ab1780) at 0.5 µg/ml
Lane 10 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) with Human Histone H3 (di methyl K27) peptide (ab1781) at 0.5 µg/ml
Lane 11 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) with Human Histone H3 (tri methyl K27) peptide (ab1782) at 0.5 µg/ml
Lysates/proteins at 0.5 µg per lane.
Secondary
All lanes : IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 17 kDa
Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (mono methyl K9) antibody - ChIP Grade (ab9045)
ab9045 staining Histone H3 (mono methyl K9) in HeLa cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Triton for 1h. The cells were then incubated with the antibody ab9045 at 0.1µg/ml and ab7291 (Mouse monoclonal to alpha Tubulin - Loading Control) used at a 1/1000 dilution overnight at +4°C. The secondary antibodies were ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed, (pseudo-colored green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594) preadsorbed, (colored red), both used at a 1/1000 dilution for 1 hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43 µM for 1hour at room temperature.
Immunoprecipitation - Anti-Histone H3 (mono methyl K9) antibody - ChIP Grade (ab9045)
Histone H3 (mono methyl K9) was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to Histone H3 (mono methyl K9) and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab9045.
Secondary: Anti-rabbit IgG VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution.
Band: 17kDa: Histone H3 (mono methyl K9).
