Rabbit anti-H3K27me3 Recombinant Monoclonal Antibody(431-118)描述别名宿主特异性反应种属应用免疫原形式浓度纯化方法类型克隆号储存/保存方法存储溶液背景说明组织特异性翻译后修饰细胞定位UniProt
| 概述 | |
| 描述 |
Modulation of chromatin structure plays an important role in the regulation of transcription in eukaryotes. The nucleosome, made up of DNA wound around eight core histone proteins (two each of H2A, H2B, H3, and H4), is the primary building block of chromatin (1). The amino-terminal tails of core histones undergo various post-translational modifications, including acetylation, phosphorylation, methylation, and ubiquitination (2-5). These modifications occur in response to various stimuli and have a direct effect on the accessibility of chromatin to transcription factors and, therefore, gene expression (6). In most species, histone H2B is primarily acetylated at Lys5, 12, 15, and 20 (4,7). Histone H3 is primarily acetylated at Lys9, 14, 18, 23, 27, and 56. Acetylation of H3 at Lys9 appears to have a dominant role in histone deposition and chromatin assembly in some organisms (2,3). Phosphorylation at Ser10, Ser28, and Thr11 of histone H3 is tightly correlated with chromosome condensation during both mitosis and meiosis (8-10). Phosphorylation at Thr3 of histone H3 is highly conserved among many species and is catalyzed by the kinase haspin. Immunostaining with phospho-specific antibodies in mammalian cells reveals mitotic phosphorylation at Thr3 of H3 in prophase and its dephosphorylation during anaphase (11).
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| 别名 |
Histone H3K27me3抗体;Histone H3 (tri methyl K27)
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| 宿主 |
Rabbit
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| 特异性 |
Histone H3K27me3 Antibody detects endogenous levels of total Histone H3K27me3.
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| 反应种属 |
Human, Mouse, Rat
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| 应用 |
WB: 1:1000, IP: 1:50, IHC-P: 1:500, ICC: 1:500, FC (Intra): 1:500
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| 免疫原 |
Synthetic peptide
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| 性能 | |
| 形式 |
Liquid
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| 浓度 |
0.5 mg/mL
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| 纯化方法 |
Protein A affinity column
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| 类型 |
Monoclonal Antibody
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| 克隆号 |
431-118
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| 储存/保存方法 |
Store at -20℃ for one year.
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| 存储溶液 |
PBS, 40% Glycerol, 0.05% BSA, 0.03% Proclin 300
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| 靶标 | |
| 背景说明 |
H3K27me3 is an important type of transliteration inhibitory translation modification. It mainly exists in the promoter of gene -dense areas and plays a role in all aspects of the biological process with the development regulatory factors in the embryo stem cells. H3K27me3 is lacking in the expression of about 50% malignant peripheral nerve sheath tumor (MPNST), and the nerve sheath tumor is positive. In pathology, it is mainly used in the differentiated diagnosis of malignant peripheral nerve sheath tumors, especially high level malignant peripheral nerve sheath tumors (lack of expression) and similar tumors.
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| 组织特异性 |
Expressed in testicular cells.
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| 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 (By similarity).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 JPent on active promoters (By similarity).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 (By similarity).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 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 Tyr-42 (H3Y41ph) by JAK2 promotes exclusion of CBX5 (HP1 alpha) from chromatin (By similarity).Ubiquitinated.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 (By similarity).
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| 细胞定位 |
Nucleus
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| UniProt |
P68431
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实验结果图
WB result of H3K27me3 Rabbit mAb Primary antibody: H3K27me3 Rabbit mAb at 1/1000 dilution Lane 1: HeLa whole cell lysate 20 ug Lane 2: HCT 116 whole cell lysate 20 ug Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution Predicted MW: 17 kDa Observed MW: 17 kDa Exposure time: 60 s
WB result of H3K27me3 Rabbit mAb Primary antibody: H3K27me3 Rabbit mAb at 1/1000 dilution Lane 1: NIH/3T3 whole cell lysate 20 ug Lane 2: mouse brain lysate 20 ug Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution Predicted MW: 17 kDa Observed MW: 17 kDa Exposure time: 120 s
WB result of H3K27me3 Rabbit mAb Primary antibody: H3K27me3 Rabbit mAb at 1/1000 dilution Lane 1: C6 whole cell lysate 20 ug Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution Predicted MW: 17 kDa Observed MW: 17 kDa Exposure time: 60 s
H3K27me3 Rabbit mAb at 1/50 dilution (1 ug) immunoprecipitating H3K27me3 in 0.4 mg HeLa whole cell lysate. Western blot was performed on the immunoprecipitate using H3K27me3 Rabbit mAb at 1/1000 dilution. Secondary antibody (HRP) for IP was used at 1/400 dilution. Lane 1: HeLa whole cell lysate 20 ug (Input) Lane 2: H3K27me3 Rabbit mAb IP in HeLa whole cell lysate Lane 3: Rabbit monoclonal IgG IP in HeLa whole cell lysate Predicted MW: 17 kDa Observed MW: 17 kDa Exposure time: 90 s
IHC shows positive staining in paraffin-embedded human cerebral cortex. Anti-H3K27me3 antibody was used at 1/500 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded rat kidney. Anti-H3K27me3 antibody was used at 1/500 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human kidney. Anti-H3K27me3 antibody was used at 1/500 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human breast cancer. Anti-H3K27me3 antibody was used at 1/500 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human lung squamous cell carcinoma. Anti-H3K27me3 antibody was used at 1/500 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded malignant peripheral nerve sheath tumor (loss of H3K27me3 expression in partial tumor cells). Anti-H3K27me3 antibody was used at 1/500 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded nerve sheath tumor. Anti-H3K27me3 antibody was used at 1/500 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded mouse cerebral cortex. Anti-H3K27me3 antibody was used at 1/500 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded mouse liver. Anti-H3K27me3 antibody was used at 1/500 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded rat cerebral cortex. Anti-H3K27me3 antibody was used at 1/500 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
ICC shows positive staining in HCT116 cells. Anti- H3K27me3 antibody was used at 1/500 dilution (Green) and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG – H&L (Alexa Fluor® 488) was used as secondary antibody at 1/1000 dilution. The cells were fixed with 100% ice-cold methanol and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue). Counterstain with tubulin (red).
Flow cytometric analysis of 4% PFA fixed 90% methanol permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling H3K27me3 antibody at 1/500 dilution (0.1 μg) / (red) compared with a Rabbit monoclonal IgG (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti – Rabbit IgG Alexa Fluor® 488 was used as the secondary antibody.
Dot blot result of H3K27me3 Rabbit mAb Lane 1: H3K27me3 peptide Lane 2: H3K27me2 peptide Lane 1: H3K27un peptide Lane 2: H3K27me1 peptide Primary antibody: H3K27me3 Rabbit mAb at 1/1000 dilution Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution Exposure time: 90 s