概述
产品名称
Histone H3 (mono methyl K4) Rabbit Polyclonal Antibody
抗体类型
Rabbit Polyclonal Antibody
免疫原
Synthetic peptide within human Histone H3 aa 1-49/136.
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
NCCIT, PC-12, MCF-7, human liver carcinoma tissue, human colon carcinoma tissue, human pancreas tissue, SHSY5Y.
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℃ or -80℃. 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
-
WB
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1:500-1:2000
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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
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., Mi L.-Z., Chruszcz M., Cymborowski M., Clines K.L., Kim Y., Minor W., Rastinejad F., Khorasanizadeh S."Double chromodomains cooperate to recognize the methylated histone H3 tail."Nature 438:1181-1185(2005)
2. "Arginine methylation of the histone H3 tail impedes effector binding."Iberg A.N., Espejo A., Cheng D., Kim D., Michaud-Levesque J., Richard S., Bedford M.T.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
别名
HIST1 cluster, H3J antibody
Histone gene cluster 1, H3 histone family, member E antibody
Histone gene cluster 1, H3G antibody
H3 histone family, member J antibody
HIST1 cluster, H3E antibody
HIST1 cluster, H3I antibody
Histone gene cluster 1, H3C antibody
FLJ92264 antibody
H3 histone family, member A antibody
H3 histone family, member B antibody
展开HIST1 cluster, H3J antibody
Histone gene cluster 1, H3 histone family, member E antibody
Histone gene cluster 1, H3G antibody
H3 histone family, member J antibody
HIST1 cluster, H3E antibody
HIST1 cluster, H3I antibody
Histone gene cluster 1, H3C antibody
FLJ92264 antibody
H3 histone family, member A antibody
H3 histone family, member B antibody
H3 histone family, member C antibody
H3 histone family, member D antibody
H3 histone family, member F antibody
H3 histone family, member H antibody
H3 histone family, member I antibody
H3 histone family, member K antibody
H3 histone family, member L antibody
H3 histone family, member T antibody
H3 histone, family 3A antibody
H3.1 antibody
H3.3A antibody
H3/a antibody
H3/b antibody
H3/c antibody
H3/d antibody
h3/f antibody
H3/h antibody
H3/i antibody
H3/j antibody
H3/k antibody
H3/l antibody
H3/t antibody
H31_HUMAN antibody
H3F1K antibody
H3F3 antibody
H3F3A antibody
H3FA antibody
H3FB antibody
H3FC antibody
H3FD antibody
H3FF antibody
H3FH antibody
H3FI antibody
H3FJ antibody
H3FK antibody
H3FL antibody
HIST1 cluster, H3A antibody
HIST1 cluster, H3B antibody
HIST1 cluster, H3C antibody
HIST1 cluster, H3D antibody
HIST1 cluster, H3F antibody
HIST1 cluster, H3G antibody
HIST1 cluster, H3H antibody
HIST1H3A antibody
HIST1H3B antibody
HIST1H3C antibody
HIST1H3D antibody
HIST1H3E antibody
HIST1H3F antibody
HIST1H3G antibody
HIST1H3H antibody
HIST1H3I antibody
HIST1H3J antibody
HIST3H3 antibody
Histone 1, H3a antibody
Histone 1, H3b antibody
Histone 1, H3c antibody
Histone 1, H3d antibody
Histone 1, H3e antibody
Histone 1, H3f antibody
Histone 1, H3g antibody
Histone 1, H3h antibody
Histone 1, H3i antibody
Histone 1, H3j antibody
histone 3, H3 antibody
histone cluster 1 H3 family member a antibody
histone cluster 1 H3 family member b antibody
histone cluster 1 H3 family member c antibody
histone cluster 1 H3 family member d antibody
histone cluster 1 H3 family member e antibody
histone cluster 1 H3 family member f antibody
histone cluster 1 H3 family member g antibody
histone cluster 1 H3 family member h antibody
histone cluster 1 H3 family member i antibody
histone cluster 1 H3 family member j antibody
Histone cluster 1, H3a antibody
Histone cluster 1, H3b antibody
Histone cluster 1, H3c antibody
Histone cluster 1, H3d antibody
Histone cluster 1, H3e antibody
Histone cluster 1, H3f antibody
Histone cluster 1, H3g antibody
Histone cluster 1, H3i antibody
Histone cluster 1, H3j antibody
Histone gene cluster 1, H3 histone family, member A antibody
Histone gene cluster 1, H3 histone family, member B antibody
Histone gene cluster 1, H3 histone family, member C antibody
Histone gene cluster 1, H3 histone family, member D antibody
Histone gene cluster 1, H3 histone family, member F antibody
Histone gene cluster 1, H3 histone family, member G antibody
Histone gene cluster 1, H3 histone family, member H antibody
Histone gene cluster 1, H3 histone family, member I antibody
Histone gene cluster 1, H3 histone family, member J antibody
Histone gene cluster 1, H3A antibody
Histone gene cluster 1, H3B antibody
Histone gene cluster 1, H3D antibody
Histone gene cluster 1, H3E antibody
Histone gene cluster 1, H3F antibody
Histone gene cluster 1, H3H antibody
Histone gene cluster 1, H3I antibody
Histone gene cluster 1, H3J antibody
Histone H 3 antibody
Histone H3.1 antibody
histone H3.1t antibody
Histone H3.2 antibody
Histone H3/a antibody
Histone H3/b antibody
Histone H3/c antibody
Histone H3/d antibody
Histone H3/f antibody
Histone H3/h antibody
Histone H3/i antibody
Histone H3/j antibody
Histone H3/k antibody
Histone H3/l antibody
Histone H3/m antibody
Histone H3/o antibody
H3K4me1 antibody
折叠图片
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☑ Cell treatment (CT)
Western blot analysis of Histone H3 (mono 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 (R1110-1, 1/500) was used 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:
A: NCCIT
B: purified 293T Histone
C: F9
D: F9 with Histone H3 peptide-unmodifed
E: F9 with Histone H3 peptide-mono methyl K4 lysates using anti-Histone H3(mono methyl K4) polyclonal antibody. -
Western blot analysis of Histone H3 (mono methyl K4) on PC-12 cell lysates with Rabbit anti-Histone H3 (mono methyl K4) antibody (R1110-1) at 1/500 dilution.
Lysates/proteins at 10 µg/Lane.
Predicted band size: 15 kDa
Observed band size: 15 kDa
Exposure time: 2 minutes;
10% 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 (R1110-1) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature. -
ICC staining of Histone H3 (mono methyl K4) in NCCIT 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 primary antibody (R1110-1, 1/100) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
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Immunocytochemistry analysis of MCF-7 cells labeling Histone H3 (mono methyl K4) with Rabbit anti-Histone H3 (mono methyl K4) antibody (R1110-1) at 1/200 dilution.
Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-Histone H3 (mono methyl K4) antibody (R1110-1) at 1/200 dilution in 2% negative goat serum 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. -
Immunohistochemical analysis of paraffin-embedded human liver carcinoma tissue using anti-Histone H3 (mono methyl K4) antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) 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 primary antibody (R1110-1, 1/200) for 30 minutes 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.
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Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue using anti-Histone H3 (mono methyl K4) antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) 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 primary antibody (R1110-1, 1/200) for 30 minutes 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.
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Immunohistochemical analysis of paraffin-embedded human pancreas tissue using anti-Histone H3 (mono methyl K4) antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) 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 primary antibody (R1110-1, 1/200) for 30 minutes 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.
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Flow cytometric analysis of Histone H3 (mono methyl K4) was done on SHSY5Y cells. The cells were fixed, permeabilized and stained with the primary antibody (R1110-1, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
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Dot blot analysis of Histone H3 (mono methyl K4) on different proteins with Rabbit anti-Histone H3 (mono methyl K4) antibody (R1110-1) 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) (positive)
Lane 3: Di-Methyl-Histone H3 (Lys4) (negative)
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: 50 seconds.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"