Anti-Histone H3 (mono methyl K36) 抗体 - ChIP Grade (ab9048)
Key features and details
- Rabbit polyclonal to Histone H3 (mono methyl K36) - ChIP Grade
- Suitable for: ICC, WB, ChIP
- Reacts with: Human
- Isotype: IgG
製品の概要
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製品名
Anti-Histone H3 (mono methyl K36) antibody - ChIP Grade
Histone H3 一次抗体 製品一覧 -
製品の詳細
Rabbit polyclonal to Histone H3 (mono methyl K36) - ChIP Grade -
由来種
Rabbit -
特異性
Specific for human Histone H3 mono methyl K36. Shows partial cross-reactivity with di-methyl K36 (please see Western Blot image). This antibody may not be suitable for experiments on yeast lysate. Although the antibody is specifically blocked using the immunising peptide, customer feedback indicates that it detects a band using S. cerevisiae K36 point mutants. We welcome further customer feedback. -
アプリケーション
適用あり: ICC, WB, ChIPmore details -
種交差性
交差種: Human
交差が予測される動物種: Mouse, Cow, Pig, Saccharomyces cerevisiae, Arabidopsis thaliana, Drosophila melanogaster, Mammals
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免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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特記事項
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
製品の特性
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製品の状態
Liquid -
保存方法
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. -
バッファー
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: 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. -
Concentration information loading... -
精製度
Immunogen affinity purified -
ポリ/モノ
ポリクローナル -
アイソタイプ
IgG -
研究分野
関連製品
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ChIP Related Products
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Compatible Secondaries
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Control Peptide
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Isotype control
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Recombinant Protein
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Related Products
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab9048の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
| アプリケーション | Abreviews | 特記事項 |
|---|---|---|
| ICC | (2) |
Use at an assay dependent concentration.
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| WB | (4) |
1/1000.
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| ChIP | (3) |
Use 2 µg for 25 µg of chromatin.
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| 特記事項 |
|---|
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ICC
Use at an assay dependent concentration. |
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WB
1/1000. |
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ChIP
Use 2 µg for 25 µg of chromatin. |
ターゲット情報
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機能
Core component of nucleosome. Nucleosomes wrap and compact DNA into chromatin, limiting DNA accessibility to the cellular machineries which require DNA as a template. Histones thereby play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling. -
配列類似性
Belongs to the histone H3 family. -
発生段階
Expressed during S phase, then expression strongly decreases as cell division slows down during the process of differentiation. -
翻訳後修飾
Acetylation is generally linked to gene activation. Acetylation on Lys-10 (H3K9ac) impairs methylation at Arg-9 (H3R8me2s). Acetylation on Lys-19 (H3K18ac) and Lys-24 (H3K24ac) favors methylation at Arg-18 (H3R17me).
Citrullination at Arg-9 (H3R8ci) and/or Arg-18 (H3R17ci) by PADI4 impairs methylation and represses transcription.
Asymmetric dimethylation at Arg-18 (H3R17me2a) by CARM1 is linked to gene activation. Symmetric dimethylation at Arg-9 (H3R8me2s) by PRMT5 is linked to gene repression. Asymmetric dimethylation at Arg-3 (H3R2me2a) by PRMT6 is linked to gene repression and is mutually exclusive with H3 Lys-5 methylation (H3K4me2 and H3K4me3). H3R2me2a is present at the 3' of genes regardless of their transcription state and is enriched on inactive promoters, while it is absent on active promoters.
Methylation at Lys-5 (H3K4me), Lys-37 (H3K36me) and Lys-80 (H3K79me) are linked to gene activation. Methylation at Lys-5 (H3K4me) facilitates subsequent acetylation of H3 and H4. Methylation at Lys-80 (H3K79me) is associated with DNA double-strand break (DSB) responses and is a specific target for TP53BP1. Methylation at Lys-10 (H3K9me) and Lys-28 (H3K27me) are linked to gene repression. Methylation at Lys-10 (H3K9me) is a specific target for HP1 proteins (CBX1, CBX3 and CBX5) and prevents subsequent phosphorylation at Ser-11 (H3S10ph) and acetylation of H3 and H4. Methylation at Lys-5 (H3K4me) and Lys-80 (H3K79me) require preliminary monoubiquitination of H2B at 'Lys-120'. Methylation at Lys-10 (H3K9me) and Lys-28 (H3K27me) are enriched in inactive X chromosome chromatin.
Phosphorylated at Thr-4 (H3T3ph) by GSG2/haspin during prophase and dephosphorylated during anaphase. Phosphorylation at Ser-11 (H3S10ph) by AURKB is crucial for chromosome condensation and cell-cycle progression during mitosis and meiosis. In addition phosphorylation at Ser-11 (H3S10ph) by RPS6KA4 and RPS6KA5 is important during interphase because it enables the transcription of genes following external stimulation, like mitogens, stress, growth factors or UV irradiation and result in the activation of genes, such as c-fos and c-jun. Phosphorylation at Ser-11 (H3S10ph), which is linked to gene activation, prevents methylation at Lys-10 (H3K9me) but facilitates acetylation of H3 and H4. Phosphorylation at Ser-11 (H3S10ph) by AURKB mediates the dissociation of HP1 proteins (CBX1, CBX3 and CBX5) from heterochromatin. Phosphorylation at Ser-11 (H3S10ph) is also an essential regulatory mechanism for neoplastic cell transformation. Phosphorylated at Ser-29 (H3S28ph) by MLTK isoform 1, RPS6KA5 or AURKB during mitosis or upon ultraviolet B irradiation. Phosphorylation at Thr-7 (H3T6ph) by PRKCBB is a specific tag for epigenetic transcriptional activation that prevents demethylation of Lys-5 (H3K4me) by LSD1/KDM1A. At centromeres, specifically phosphorylated at Thr-12 (H3T11ph) from prophase to early anaphase, by DAPK3 and PKN1. Phosphorylation at Thr-12 (H3T11ph) by PKN1 is a specific tag for epigenetic transcriptional activation that promotes demethylation of Lys-10 (H3K9me) by KDM4C/JMJD2C. Phosphorylation at Tyr-42 (H3Y41ph) by JAK2 promotes exclusion of CBX5 (HP1 alpha) from chromatin.
Monoubiquitinated by RAG1 in lymphoid cells, monoubiquitination is required for V(D)J recombination (By similarity). Ubiquitinated by the CUL4-DDB-RBX1 complex in response to ultraviolet irradiation. This may weaken the interaction between histones and DNA and facilitate DNA accessibility to repair proteins. -
細胞内局在
Nucleus. Chromosome. - Information by UniProt
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参照データベース
- 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
see all -
別名
- H3 histone family member E pseudogene antibody
- H3 histone family, member A antibody
- H3/A antibody
see all
画像
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ChIP - Anti-Histone H3 (mono methyl K36) antibody - ChIP Grade (ab9048)
Chromatin was prepared from HeLa cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 2µg of ab9048 (blue), and 20µl 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 - Anti-Histone H3 (mono methyl K36) antibody - ChIP Grade (ab9048)All lanes : Anti-Histone H3 (mono methyl K36) antibody - ChIP Grade (ab9048) at 1/500 dilution
Lane 1 : Histone prep
Lane 2 : Histone prep with Human Histone H3 (mono methyl K36) peptide (ab1783) at 1 µg/ml
Lane 3 : Histone prep withHuman Histone H3 (di methyl K36) peptide (ab1784) at 1 µg/ml
Lane 4 : Histone prep withHuman Histone H3 (tri methyl K36) peptide (ab1785) at 1 µg/ml
Lane 5 : Histone prep withHuman Histone H3 (unmodified ) peptide (ab2623) at 1 µg/ml
Lane 6 : Histone prep withHuman Histone H3 (mono methyl K4) peptide (ab1340) at 1 µg/ml
Lysates/proteins at 0.5 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab6721) at 1/5000 dilutionSecondary ab: Alexa Fluor 680 Goat anti-rabbit IgG
0.5µ Primary antibody:
Lane 1: ab9048 (Histone H3 Mono Methyl K36) 1/500
Lane 2: ab9048 (Histone H3 Mono Methyl K36) 1/500 + ab1783 (ab9048) (Histone H3 Mono Methyl K36) peptide 1µ
Lane 3: ab9048 (Histone H3 Mono Methyl K36) 1/500 + ab1794 (ab9049) (Histone H3 Di Methyl K36) peptide 1µ
Lane 4: ab9048 (Histone H3 Mono Methyl K36) 1/500 + ab1785 (ab9050) (Histone H3 Tri Methyl K36) peptide 1µ
Lane 5: ab9048 (Histone H3 Mono Methyl K36) 1/500 + ab2623 (Histone H3 (23-34) – unmodified) peptide 1µ
Lane 6: ab9048 (Histone H3 Mono Methyl K36) 1/500 + ab1340 (ab8895) (Histone H3 Mono methyl K4) peptide 1µ ab9048 specifically recognise
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Immunocytochemistry - Anti-Histone H3 (mono methyl K36) antibody - ChIP Grade (ab9048)ab9048 staining Histone H3 (mono methyl K36) in HeLa cells. The cells were fixed with 100% methanol (5 min) at room temperature, 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-Tween for 1h. The cells were then incubated with the antibody ab9048 at 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.
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Immunocytochemistry - Anti-Histone H3 (mono methyl K36) antibody - ChIP Grade (ab9048)This image was submitted as part of a review by Kirk McManus, University of ColumbiaStaining of interphase nuclei of Hela cells with ab9048 (green) at a working dilution of 1/500. The DNA is stained with DAPI. ab9048 appears to be more associated with heterochromatin (DAPI intense regions) than euchromatin (DAPI less intense regions).
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Western blot - Anti-Histone H3 (mono methyl K36) antibody - ChIP Grade (ab9048)Image courtesy of an anonymous Abreview.All lanes : Anti-Histone H3 (mono methyl K36) antibody - ChIP Grade (ab9048) at 1/1400 dilution
All lanes : Whole cell lysate prepared from Drosophila BG3 cells
Lysates/proteins at 500000 cells per lane.
Secondary
All lanes : HRP donkey anti-rabbit monoclonal at 1/20000 dilution
Developed using the ECL technique.
Observed band size: 17,42 kDa why is the actual band size different from the predicted?
Exposure time: 30 seconds
プロトコール
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
データシートおよび資料
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SDS download
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Datasheet download
参考文献 (103)
ab9048 は 103 報の論文で使用されています。
- Chen F et al. Histone Lysine Methyltransferase SETD2 Regulates Coronary Vascular Development in Embryonic Mouse Hearts. Front Cell Dev Biol 9:651655 (2021). PubMed: 33898448
- Shao R et al. H3K36 methyltransferase NSD1 regulates chondrocyte differentiation for skeletal development and fracture repair. Bone Res 9:30 (2021). PubMed: 34099628
- Lerner AM et al. An optogenetic switch for the Set2 methyltransferase provides evidence for transcription-dependent and -independent dynamics of H3K36 methylation. Genome Res 30:1605-1617 (2020). PubMed: 33020206
- DiFiore JV et al. Unique and Shared Roles for Histone H3K36 Methylation States in Transcription Regulation Functions. Cell Rep 31:107751 (2020). PubMed: 32521276
- Liu DJ et al. setd2 knockout zebrafish is viable and fertile: differential and developmental stress-related requirements for Setd2 and histone H3K36 trimethylation in different vertebrate animals. Cell Discov 6:72 (2020). PubMed: 33088589