概述
产品名称
Histone H3 (di methyl K4) Rabbit Polyclonal Antibody
抗体类型
Rabbit Polyclonal Antibody
免疫原
Synthetic peptide of the N terminal residues of Human Di-mehtyl-Histone H3(Lys4).
种属反应性
Human, Mouse, Rat
验证应用
WB, IF-Cell, IHC-P, FC, Dot Blot
分子量
Predicted band size: 15 kDa
阳性对照
Human liver tissue, Histone, F9, F9 treated with Histone H3 peptide-unmodifed, F9 treated with Histone H3 peptide-di-methyl K4, Ags, Hela, HepG2, rat testis tissue, mouse testis tissue.
偶联
unconjugated
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℃. Avoid repeated freeze / thaw cycles.
存储缓冲液
1*PBS (pH7.4), 0.2% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
亚型
IgG
纯化方式
Immunogen affinity purified.
应用稀释度
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WB
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1:1,000
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IF-Cell
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1:50-1:200
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IHC-P
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1:50-1:200
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FC
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1:50-1:100
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Dot Blot
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1:1,000
发表文章中的应用
靶点
功能
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. 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. 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.
背景文献
1. Flanagan J.F et al. Double chromodomains cooperate to recognize the methylated histone H3 tail. Nature 438:1181-1185 (2005).
2. Iberg A.N et al. Arginine methylation of the histone H3 tail impedes effector binding. J Biol Chem 283:3006-3010 (2008).
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.
UNIPROT #
图片
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Western blot analysis of Histone H3 (di methyl K4) on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody was used at a 1:1,000 dilution in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: Human liver tissue lysate, untreated
Lane 2: Histone lysate, purified from 293T
Lane 3: F9 cell lysate, untreated
Lane 4: F9 cell lysate, treated with Histone H3 peptide-unmodifed
Lane 5: F9 cell lysate, treated with Histone H3 peptide-di-methyl K4 -
ICC staining Histone H3 (di methyl K4) in Ags cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the antibody (R1110-3) at a dilution of 1:200 for 1 hour at room temperature, washed with PBS. Alexa Fluorc™ 488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/100 dilution. The nuclear counter stain is DAPI (blue).
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ICC staining Histone H3 (di methyl K4) in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the antibody (R1110-3) at a dilution of 1:200 for 1 hour at room temperature, washed with PBS. Alexa Fluorc™ 488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/100 dilution. The nuclear counter stain is DAPI (blue).
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ICC staining Histone H3 (di methyl K4) in HepG2 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the antibody (R1110-3) at a dilution of 1:200 for 1 hour at room temperature, washed with PBS. Alexa Fluorc™ 488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/100 dilution. The nuclear counter stain is DAPI (blue).
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Immunohistochemical analysis of paraffin-embedded rat testis tissue using anti-Histone H3 (di methyl K4) antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the antibody (R1110-3) at 1/200 dilution, for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chrogen. Counter stained with hematoxylin and mounted with DPX.
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Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-Histone H3 (di methyl K4) antibody (R1110-3) at 1/1,000 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 (R1110-3) at 1/1,000 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 testis tissue using anti-Histone H3 (di methyl K4) antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the antibody (R1110-3) at 1/200 dilution, for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chrogen. Counter stained with hematoxylin and mounted with DPX.
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Flow cytometric analysis of Histone H3 (di methyl K4) was done on Hela cells. The cells were fixed, permeabilized and stained with Histone H3 (di methyl K4) antibody at 1/100 dilution (blue) compared with an unlabelled control (cells without incubation with primary antibody; red). After incubation of the primary antibody on room temperature for an hour, the cells was stained with a Alexa Fluor™ 488-conjugated goat anti-rabbit IgG Secondary antibody at 1/500 dilution for 30 minutes.
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Dot blot analysis of Histone H3 (di methyl K4) on different proteins with Rabbit anti-Histone H3 (di methyl K4) antibody (R1110-3) at 1/1,000 dilution. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution for 1 hour at room temperature.
Lane 1: Unmodified Histone H3 (negative)
Lane 2: Mono-Methyl-Histone H3 (Lys4) (negative)
Lane 3: Di-Methyl-Histone H3 (Lys4) (positive)
Lane 4: Tri-Methyl-Histone H3 (Lys4) (negative)
Lane 5: Acetyl-Histone H3 (Lys4) (negative)
Proteins loading: 100ng, 25ng, 5ng;
Blocking and dilution buffer: 5% NFDM/TBST;
Exposure time: 1 minute 30 seconds.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
引文
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Sanguinarine, identified as a natural alkaloid LSD1 inhibitor, suppresses lung cancer cell growth and migration
Author: Qin, T. T., Li, Z. H., Li, L. X., Du, K., Yang, J. G., Zhang, Z. Q., Wu, X. X., & Ma, J. L.
PMID: 35949313
应用: WB
反应种属: Human
发表时间: 2022 Jun
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Citation
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Targeting the LSD1-G9a-ER Stress Pathway as a Novel Therapeutic Strategy for Esophageal Squamous Cell Carcinoma
Author: Wang, H., Song, Z., Xie, E., Chen, J., Tang, B., Wang, F., & Min, J.
PMID: 35707047
应用: IHC
反应种属: Mouse
发表时间: 2022 Jun
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Citation