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宿主来源
Rabbit抗原名称
Histone H3 (tri methyl K27)分子别名
H3K27me3细胞定位
NucleusAccession
P68431克隆号
S-4098抗体类型
Recombinant mAb抗体同种型
IgG翻译后修饰类型
甲基化反应种属 ?
Hu, Ms, Rt阳性样本
HeLa, NIH/3T3纯化方式
Protein A浓度
1 mg/ml标记
Unconjugated性状
Liquid缓冲体系
PBS, 40% Glycerol, 0.05% BSA, 0.03% Proclin 300
储存条件
12 months from date of receipt / reconstitution, -20 °C as supplied
应用
ChIP
IHC-P ?
ICC ?
WB
稀释度
应用 稀释度 推荐种属 WB 1:1000 Hu, Ms IHC-P 1:100 Hu, Ms, Rt ICC 1:500 Hu, Ms ChIP 1:20-1:50 Hu
Histone H3 lysine 27 trimethylation (abbreviated as H3K27me3) is a classic transcription-repressive epigenetic mark, catalyzed by the core catalytic subunits EZH1 or EZH2 of Polycomb Repressive Complex 2 (PRC2). This modification serves as the central executor of PRC2-mediated gene silencing. Its primary function is to recruit and stabilize effector proteins, such as the PRC1 complex, collectively promoting the formation of compact, higher-order chromatin structures to stably and long-term suppress the expression of specific genes at the epigenetic level. H3K27me3 plays a crucial role in development, extensively participating in key biological processes including cell fate determination, maintenance of stem cell pluripotency, somatic cell reprogramming, and X-chromosome inactivation. By precisely establishing and maintaining a "gene expression blueprint," it ensures the correct identity of cells. However, when this powerful silencing function becomes dysregulated, it can lead to severe consequences. For instance, gain-of-function mutations or aberrant expression of genes encoding PRC2 components (such as EZH2) result in disrupted H3K27me3 levels, a phenomenon widely observed in various malignancies including lymphoma, prostate cancer, sarcoma, and glioma, making it a key driver of tumorigenesis. Notably, its function is often antagonistic to the activating mark H3K4me3, and together they can form "bivalent chromatin domains," which co-mark key developmental regulator genes in embryonic stem cells that are poised for activation upon differentiation. Therefore, H3K27me3 is not only a central molecule in developmental biology and disease research but has also become an important target for the development of current epigenetic drugs, such as EZH2 inhibitors.
免疫印迹
WB result of Histone H3 (tri methyl K27) Recombinant Rabbit mAb
Primary antibody: Histone H3 (tri methyl K27) Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: HeLa whole cell lysate 20 µg
Secondary antibody: Goat Anti- rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 15 kDa
Observed MW: 17 kDa
This blot was developed with high sensitivity substrateWB result of Histone H3 (tri methyl K27) Recombinant Rabbit mAb
Primary antibody: Histone H3 (tri methyl K27) Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: NIH/3T3 whole cell lysate 20 µg
Secondary antibody: Goat Anti- rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 15 kDa
Observed MW: 17 kDa
This blot was developed with high sensitivity substrate
免疫组化
IHC shows positive staining in paraffin-embedded human tonsil. Anti-Histone H3 (tri methyl K27) antibody was used at 1/100 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 spleen. Anti-Histone H3 (tri methyl K27) antibody was used at 1/100 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-Histone H3 (tri methyl K27) antibody was used at 1/100 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.
免疫细胞化学
ICC shows positive staining in HeLa cells. Anti-Histone H3 (tri methyl K27) 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).
ICC shows positive staining in NIH/3T3 cells. Anti-Histone H3 (tri methyl K27) 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).
ChIP
Chromatin immunoprecipitation (ChIP) was performed on HeLa cells cross-linked with 1% formaldehyde for 10 min, then chromatin was fragmented by sonication. Parallel reactions used Histone H3 (tri methyl K27) Recombinant Rabbit mAb (S-4098) and Rabbit mAb IgG Isotype Control (SDT-R173) at 1:50 for immunoprecipitation.
Post-immunoprecipitation, both samples were washed, eluted, and cross-links reversed. Purified DNA was analyzed by qPCR.
qPCR showed the enrichment of RPL30, GAPDH, MYOD1, AFM and SAT-α in
Histone H3 (tri methyl K27) Recombinant Rabbit mAb (S-4098) -immunoprecipitated
sample.
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