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Rabbit抗原名称
H3K27me3分子别名
H3K27me3细胞定位
NucleusAccession
P68431克隆号
SDT-431-118抗体类型
Recombinant mAb抗体同种型
IgG翻译后修饰类型
甲基化应用
ChIP ,ICFCM ,IHC-P ? ,ICC ? ,WB ,IP反应种属 ?
Hu, Ms, Rt预测反应种属
(反应种属缩写表)SeUr, Fu, Hm, Pl, Dr纯化方式
Protein A浓度
0.5 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 | 稀释度 |
|---|---|
| Dot Blot | 1:1000 |
| WB | 1:1000 |
| IP | 1:50 |
| IHC | 1:500 |
| ICC | 1:500 |
| ICFCM | 1:500 |
| ChIP | 1:20-1:50 |
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.
免疫印迹
WB result of H3K27me3 Rabbit mAb
Primary antibody: H3K27me3 Rabbit mAb at 1/1000 dilution
Lane 1: HeLa whole cell lysate 20 µg
Lane 2: HCT 116 whole cell lysate 20 µg
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 sWB result of H3K27me3 Rabbit mAb
Primary antibody: H3K27me3 Rabbit mAb at 1/1000 dilution
Lane 1: NIH/3T3 whole cell lysate 20 µg
Lane 2: mouse brain lysate 20 µg
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 sWB result of H3K27me3 Rabbit mAb
Primary antibody: H3K27me3 Rabbit mAb at 1/1000 dilution
Lane 1: C6 whole cell lysate 20 µg
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
流式分析
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.
免疫沉淀
H3K27me3 Rabbit mAb at 1/50 dilution (1 µg) 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 µg (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
斑点杂交
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
免疫组化
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 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.
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.
免疫细胞化学
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).
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 (SDT-431-118) 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 (%input: immunoprecipitated DNA/input DNA)
showed the enrichment of RPL30, GAPDH, MYOD1,
AFM, SAT-α and SAT-2 in Histone H3 (tri methyl K27) Recombinant
Rabbit mAb (SDT-431-118)-immunoprecipitated sample.
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