概述
产品名称
Phospho-Histone H3 (S10) Recombinant Rabbit Monoclonal Antibody [SA31-01]
抗体类型
Recombinant Rabbit monoclonal Antibody
免疫原
Synthetic phospho-peptide corresponding to residues surrounding Ser10 of human Histone H3.
种属反应性
Human, Mouse, Rat
验证应用
WB, IF-Cell, IF-Tissue, IHC-P, IP, FC, ChIP
分子量
Predicted band size: 15 kDa
阳性对照
HeLa treated with 50nM Calyculin A for 30 minutes whole cell lysate, NIH/3T3 treated with 100nM Calyculin A for 30 minutes whole cell lysate, C6 treated with 100nM Calyculin A for 30 minutes whole cell lysate, HeLa, human kidney tissue, mouse kidney tissue, rat kidney tissue.
偶联
unconjugated
克隆号
SA31-01
RRID
产品特性
形态
Liquid
浓度
1ug/ul
存放说明
Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
存储缓冲液
1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
亚型
IgG
纯化方式
Protein A affinity purified.
应用稀释度
-
WB
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1:2,000-1:10,000
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IF-Cell
-
1:5,000
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IF-Tissue
-
1:50
-
IHC-P
-
1:100
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IP
-
Use at an assay dependent concentration.
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FC
-
1:1,000
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ChIP
-
Use 0.5~2 μg for 25 μg of chromatin.
发表文章中的应用
WB | See 2 publications below |
IF-cell | See 1 publications below |
发表文章中的种属
靶点
功能
Histone H3 is one of the five main histones involved in the structure of chromatin in eukaryotic cells. Featuring a main globular domain and a long N-terminal tail, H3 is involved with the structure of the nucleosomes of the 'beads on a string' structure. Histone proteins are highly post-translationally modified however Histone H3 is the most extensively modified of the five histones. The term "Histone H3" alone is purposely ambiguous in that it does not distinguish between sequence variants or modification state. Histone H3 is an important protein in the emerging field of epigenetics, where its sequence variants and variable modification states are thought to play a role in the dynamic and long term regulation of genes.
背景文献
1. Hammond SL et al. Mitotic phosphorylation of histone H3 threonine 80. Cell Cycle 13:440-52 (2014).
2. Martin HL et al. High-content, high-throughput screening for the identification of cytotoxic compounds based on cell morphology and cell proliferation markers. PLoS One 9:e88338 (2014).
序列相似性
Belongs to the histone H3 family.
翻译后修饰
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). Acetylation at Lys-123 (H3K122ac) by EP300/p300 plays a central role in chromatin structure: localizes at the surface of the histone octamer and stimulates transcription, possibly by promoting nucleosome instability.; 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. Monomethylation at Lys-57 (H3K56me1) by EHMT2/G9A in G1 phase promotes interaction with PCNA and is required for DNA replication.; Phosphorylated at Thr-4 (H3T3ph) by 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 MAP3K20 isoform 1, RPS6KA5 or AURKB during mitosis or upon ultraviolet B irradiation. Phosphorylation at Thr-7 (H3T6ph) by PRKCB 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 Thr-12 (H3T11ph) by chromatin-associated CHEK1 regulates the transcription of cell cycle regulatory genes by modulating acetylation of Lys-10 (H3K9ac). 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.; Lysine deamination at Lys-5 (H3K4all) to form allysine is mediated by LOXL2. Allysine formation by LOXL2 only takes place on H3K4me3 and results in gene repression.; Crotonylation (Kcr) is specifically present in male germ cells and marks testis-specific genes in post-meiotic cells, including X-linked genes that escape sex chromosome inactivation in haploid cells. Crotonylation marks active promoters and enhancers and confers resistance to transcriptional repressors. It is also associated with post-meiotically activated genes on autosomes.; Butyrylation of histones marks active promoters and competes with histone acetylation. It is present during late spermatogenesis.; Succinylation at Lys-80 (H3K79succ) by KAT2A takes place with a maximum frequency around the transcription start sites of genes. It gives a specific tag for epigenetic transcription activation. Desuccinylation at Lys-123 (H3K122succ) by SIRT7 in response to DNA damage promotes chromatin condensation and double-strand breaks (DSBs) repair.; Serine ADP-ribosylation constitutes the primary form of ADP-ribosylation of proteins in response to DNA damage. Serine ADP-ribosylation at Ser-11 (H3S10ADPr) is mutually exclusive with phosphorylation at Ser-11 (H3S10ph) and impairs acetylation at Lys-10 (H3K9ac).
亚细胞定位
Nucleus, Chromosome
UNIPROT #
别名
H3 3 like sequence MH921 antibody
H3 3A antibody
H3 a antibody
H3 b antibody
H3 c antibody
H3 d antibody
H3 f antibody
H3 h antibody
H3 histone family member E pseudogene antibody
H3 i antibody
展开H3 3 like sequence MH921 antibody
H3 3A antibody
H3 a antibody
H3 b antibody
H3 c antibody
H3 d antibody
H3 f antibody
H3 h antibody
H3 histone family member E pseudogene antibody
H3 i antibody
H3 j antibody
H3 k antibody
H3 l antibody
H33_HUMAN antibody
H3F3 antibody
H3f3b antibody
Histone H3 3 pseudogene antibody
Histone H3.3 antibody
折叠图片
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☑ Cell treatment (CT)
Western blot analysis of Phospho-Histone H3 (S10) on different lysates with Rabbit anti-Phospho-Histone H3 (S10) antibody (ET1601-30) at 1/5,000 dilution.
Lane 1: HeLa whole cell lysate (20 µg/Lane)
Lane 2: HeLa treated with 50nM Calyculin A for 30 minutes whole cell lysate (20 µg/Lane)
Lane 3: NIH/3T3 whole cell lysate (20 µg/Lane)
Lane 4: NIH/3T3 treated with 100nM Calyculin A for 30 minutes whole cell lysate (20 µg/Lane)
Lane 5: C6 whole cell lysate (20 µg/Lane)
Lane 6: C6 treated with 100nM Calyculin A for 30 minutes whole cell lysate (20 µg/Lane)
Predicted band size: 15 kDa
Observed band size: 15 kDa
Exposure time: 10 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1601-30) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:50,000 dilution was used for 1 hour at room temperature. -
Immunocytochemistry analysis of HeLa cells labeling Phospho-Histone H3 (S10) with Rabbit anti-Phospho-Histone H3 (S10) antibody (ET1601-30) at 1/5,000 dilution and competitor's antibody at 1/3,000 dilution.
Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Phospho-Histone H3 (S10) antibody (ET1601-30) at 1/5,000 dilution and competitor's antibody at 1/3,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
☑ Cell treatment (CT)
Western blot analysis of Phospho-Histone H3 (S10) on different lysates with Rabbit anti-Phospho-Histone H3 (S10) antibody (ET1601-30) at 1/2,000 dilution and competitor's antibody at 1/1,000 dilution.
Lane 1: HeLa whole cell lysate
Lane 2: HeLa treated with 50nM Calyculin A for 30 minutes whole cell lysate
Lane 3: C6 whole cell lysate
Lane 4: C6 treated with 100ng/mL Calyculin A for 1 hour whole cell lysate
Lane 5: NIH/3T3 whole cell lysate
Lane 6: NIH/3T3 starved for 4 hours, then treated with 100nM Calyculin A for 30 minutes whole cell lysate
Lysates/proteins at 15 µg/Lane.
Predicted band size: 15 kDa
Observed band size: 15 kDa
Exposure time: 15 seconds;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1601-30) at 1/2,000 dilution and competitor's antibody at 1/1,000 dilution were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-Phospho-Histone H3 (S10) antibody (ET1601-30) at 1/100 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1601-30) at 1/100 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-Phospho-Histone H3 (S10) antibody (ET1601-30) at 1/100 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1601-30) at 1/100 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-Phospho-Histone H3 (S10) antibody (ET1601-30) at 1/100 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1601-30) at 1/100 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
☑ Cell treatment (CT)
Flow cytometric analysis of HeLa untreated cells and HeLa treated with 100nM Calyculin A for 30 minutes cells labeling Phospho-Histone H3 (S10).
Cells were fixed and permeabilized. Then stained with the primary antibody (ET1601-30, 1μg/mL) (HeLa untreated,blue; HeLa treated, red) compared with Rabbit IgG Isotype Control (grey). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). -
☑ Cell treatment (CT)
Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells treated with 100ng/mL Nocodazole for 18 hours and either Phospho-Histone H3 (S10) (ET1601-30) or Normal Rabbit IgG according to the ChIP protocol. The enriched DNA was quantified by real-time PCR using indicated primers. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
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引文
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Chaetocin exhibits anticancer effects in esophageal squamous cell carcinoma via activation of Hippo pathway
Author:
PMID: 37319316
应用: WB
反应种属: Human
发表时间: 2023 Jun
-
Citation
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Biomimetic and NOS‐Responsive Nanomotor Deeply Delivery a Combination of MSC‐EV and Mitochondrial ROS Scavenger and Promote Heart Repair and Regeneration
Author:
PMID: 37282826
应用: IF-cell
反应种属: Rat
发表时间: 2023 Jul
-
Citation
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CENPM upregulation by E5 oncoprotein of human papillomavirus promotes radiosensitivity in head and neck squamous cell carcinoma
Author:
PMID: 35462155
应用: WB
反应种属: Human
发表时间: 2022 Jun
-
Citation