Anti-Histone H3 (phospho S28) 抗体 [HTA28] (ab10543)
![Western blot - Anti-Histone H3 (phospho S28) antibody [HTA28] (ab10543)](https://www.abcam.co.jp/ps/products/10/ab10543/Images/ab10543-396058-anti-histone-h3-phospho-s28-antibody-hta28-western-blot.jpg "Western blot - Anti-Histone H3 (phospho S28) antibody [HTA28] (ab10543)")
Key features and details
- Rat monoclonal [HTA28] to Histone H3 (phospho S28)
- Suitable for: WB, ICC
- Reacts with: Human
- Isotype: IgG2a
製品の概要
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製品名
Anti-Histone H3 (phospho S28) antibody [HTA28]
Histone H3 一次抗体 製品一覧 -
製品の詳細
Rat monoclonal [HTA28] to Histone H3 (phospho S28) -
由来種
Rat -
アプリケーション
適用あり: WB, ICCmore details -
種交差性
交差種: Human
交差が予測される動物種: Mouse, Hamster, Cow
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免疫原
Synthetic peptide corresponding to Human Histone H3 aa 23-35 (phospho S28) conjugated to Keyhole Limpet Haemocyanin (KLH).
Sequence:KAARKSA PATGGV
Database link: P68431 -
ポジティブ･コントロール
- WB: HeLa (treated with Nocodazole) whole cell extract. ICC: HeLa cells.
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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: 0.097% Sodium azide
Constituents: 0.0268% PBS, 1% BSA -
Concentration information loading... -
精製度
Immunogen affinity purified -
特記事項（精製）
Purified from culture supernatant of hybridoma cells grown in a bioreactor -
ポリ/モノ
モノクローナル -
クローン名
HTA28 -
アイソタイプ
IgG2a -
研究分野
関連製品
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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ab10543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 |
|---|---|---|
| WB | (1) |
Use a concentration of 0.5 - 2 µg/ml. Detects a band of approximately 15 kDa.
We advise you to centrifuge this product vial before use. |
| ICC | (2) |
Use a concentration of 5 µg/ml.
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| 特記事項 |
|---|
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WB
Use a concentration of 0.5 - 2 µg/ml. Detects a band of approximately 15 kDa. We advise you to centrifuge this product vial before use. |
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ICC
Use a concentration of 5 µg/ml. |
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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Western blot - Anti-Histone H3 (phospho S28) antibody [HTA28] (ab10543)Lane 1 : Anti-Histone H3 (phospho S28) antibody [HTA28] (ab10543) at 0.5 µg/ml
Lane 2 : Anti-Histone H3 (phospho S28) antibody [HTA28] (ab10543) at 1 µg/ml
Lane 3 : Anti-Histone H3 (phospho S28) antibody [HTA28] (ab10543) at 2 µg/ml
All lanes : HeLa (treated with Nocodazole) whole cell extract
Secondary
All lanes : Goat Anti-Rat IgG-peroxidase conjugate
Developed using the ECL technique. -
![Immunocytochemistry - Anti-Histone H3 (phospho S28) antibody [HTA28] (ab10543)](https://www.abcam.co.jp/ps/products/10/ab10543/Images/ab10543-396057-anti-histone-h3-phospho-s28-antibody-hta28-immunocytochemistry-hela-human.jpg)
Immunocytochemistry - Anti-Histone H3 (phospho S28) antibody [HTA28] (ab10543)Immunocytochemical analysis of HeLa cells fixed and permeabilized with methanol followed by methanol:acetone. Histone H3 (phospho S28) was labeled with ab10543 at 5 μg/mL in ICC/IF (Green). The secondary antobody was a Goat Anti-Rat IgG-FITC conjugate. The nuclear counerstain was DAPI (Blue).
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![Immunocytochemistry - Anti-Histone H3 (phospho S28) antibody [HTA28] (ab10543)](https://www.abcam.co.jp/ps/products/10/ab10543/Images/ab10543-276888-anti-histone-h3-phospho-s28-antibody-hta28-immunofluorescence.jpg)
Immunocytochemistry - Anti-Histone H3 (phospho S28) antibody [HTA28] (ab10543)This image is courtesy of an anonymous Abreviewab10543 staining Histone H3 (phospho S28) in Human neural progenitor cells from iPS cell line by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.1% Triton and blocked with 1% serum, 0.1% BSA in PBS for 30 minutes at room temperature. Samples were incubated with primary antibody (2ug/ml in blocking buffer) for 16 hours at 4°C. An Alexa Fluor® 488-conjugated Goat anti-rat IgG2a polyclonal was used as the secondary antibody (1/500). Total cells were stained using DAPI (blue)
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![Western blot - Anti-Histone H3 (phospho S28) antibody [HTA28] (ab10543)](https://www.abcam.co.jp/ps/products/10/ab10543/Images/ab10543-231405-anti-histone-h3-phospho-s28-antibody-hta28-western-blot.jpg)
Western blot - Anti-Histone H3 (phospho S28) antibody [HTA28] (ab10543)All lanes : Anti-Histone H3 (phospho S28) antibody [HTA28] (ab10543) at 1 µg/ml
Lane 1 : HeLa Histone Preparation Nuclear Lysate - Colcemid-treated
Lane 2 : HeLa Histone Preparation Nuclear Lysate - Colcemid-treated withHuman Histone H3 (unmodified ) peptide (ab2623) at 0.5 µg/ml
Lane 3 : HeLa Histone Preparation Nuclear Lysate - Colcemid-treated with Human Histone H3 (phospho S28) peptide (ab5499) at 0.5 µg/ml
Lane 4 : HeLa Histone Preparation Nuclear Lysate - Colcemid-treated withHuman Histone H3 (phospho S10) peptide (ab11477) at 0.5 µg/ml
Lysates/proteins at 2.5 µg per lane.
Secondary
All lanes : Peroxidase Conjugated AffiniPure Rabbit Anti-Rat IgG (H+L) at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Observed band size: 17 kDa why is the actual band size different from the predicted?
Exposure time: 20 minutesThis blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab10543 overnight at 4°C. Antibody binding was detected using an anti-mouse antibody conjugated to HRP, and visualised using ECL development solution.
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![Immunocytochemistry - Anti-Histone H3 (phospho S28) antibody [HTA28] (ab10543)](https://www.abcam.co.jp/ps/products/10/ab10543/Images/ab10543-276889-anti-histone-h3-phospho-s28-antibody-hta28-immunofluorescence.jpg)
Immunocytochemistry - Anti-Histone H3 (phospho S28) antibody [HTA28] (ab10543)This image is courtesy of an anonymous Abreviewab10543 staining Histone H3 (phospho S28) in Mouse neural stem cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde and blocked with 4% serum for 1 hour at 20°C. Samples were incubated with primary antibody (1/500 in PBS + 4% serum) for 16 hours at 4°C. An Alexa Fluor® 546-conjugated anti-rat polyclonal was used as the secondary antibody (1/500). DAPI is stainded blue
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![Immunocytochemistry - Anti-Histone H3 (phospho S28) antibody [HTA28] (ab10543)](https://www.abcam.co.jp/ps/products/10/ab10543/Images/ab10543-207613-anti-histone-h3-phospho-s28-antibody-hta28-immunofluorescence.jpg)
Immunocytochemistry - Anti-Histone H3 (phospho S28) antibody [HTA28] (ab10543)This image is courtesy of an Abreview submitted by Dr Kirk McManusab10543 staining HeLa cells by ICC/IF. The cells were fixed with paraformaldehyde and permeabilized with 0.5% Triton X100 in PBS. The cells were then stained with ab10543 at 1/1000 in PBS for 1h at 22°C. A goat anti-rat Alexa Fluro 488 (ab150157) at 1/200 was used as the secondary antibody. Nuclei are stained in red with DAPI. The antibody produces the expected mitotic-associated staining pattern and is extremely strong. MeOH fixed samples were also evaluated and produced a similar, strong staining pattern in mitotic cells.
プロトコール
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 (88)
ab10543 は 88 報の論文で使用されています。
- Hilbert L et al. Transcription organizes euchromatin via microphase separation. Nat Commun 12:1360 (2021). PubMed: 33649325
- Li D et al. Pathogenic variants in SMARCA5, a chromatin remodeler, cause a range of syndromic neurodevelopmental features. Sci Adv 7:N/A (2021). PubMed: 33980485
- Wilmerding A et al. HoxB genes regulate neuronal delamination in the trunk neural tube by controlling the expression of Lzts1. Development 148:N/A (2021). PubMed: 33472847
- Duboc V et al. Tbx4 function during hindlimb development reveals a mechanism that explains the origins of proximal limb defects. Development 148:N/A (2021). PubMed: 34423345
- Wilmerding A et al. HOXB8 Counteracts MAPK/ERK Oncogenic Signaling in a Chicken Embryo Model of Neoplasia. Int J Mol Sci 22:N/A (2021). PubMed: 34445617