Blank control (Black line):Molt4 (Black).
Primary Antibody (green line): Rabbit Anti-phospho-Dnmt1(Ser154) antibody (bs-5294R)
Dilution: 1μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody (white blue line): Goat anti-rabbit IgG-AF647
Dilution: 1μg /test.
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 90% ice-cold methanol for 20 min at room temperature. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.
- ￥1180 - 2800
- Bioss
- bs-5294R
- 2025年10月24日
- 产品信息以Bioss网站为准
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50ul/100ul/200ul
| 规格： | 50ul | 产品价格： | ￥1180.0 |
|---|---|---|---|
| 规格： | 100ul | 产品价格： | ￥1980.0 |
| 规格： | 200ul | 产品价格： | ￥2800.0 |
| 产品编号 | bs-5294R |
| 英文名称 | phospho-Dnmt1 (Ser154) Rabbit pAb |
| 中文名称 | 磷酸化DNA甲基转移酶1抗体 |
| 英文别名 | Dnmt1(phospho S154); Dnmt1(phospho Ser154); p-Dnmt1(Ser154); AIM; CXXC finger protein 9; CXXC-type zinc finger protein 9; CXXC9; DNA(cytosine 5 ) methyltransferase 1; DNA(cytosine-5)-methyltransferase 1; DNA methyltransferase 1; DNA methyltransferase HsaI; DNA methyltransferase M.HsaI.; DNA MTase; DNA MTase HsaI; DNMT 1; DNMT; Dnmt1; Dnmt1; DNMT1_HUMAN; Dnmt1o; FLJ16293; HSN1E; M.HsaI; MCMT; Met1; MGC104992; mMmul; MommeD2. |
| 产品应用 | IHC-P=1:100-500, IHC-F=1:100-500, IF=1:100-500, Flow-Cyt=1ug/Test Not yet tested in other applications. |
| 交叉反应 | Human (Mouse, Dog, Pig, Cow, Sheep) |
| 抗体来源 | Rabbit |
| 免疫原 | KLH conjugated Synthesised phosphopeptide derived from human Dnmt1 around the phosphorylation site of Ser153 |
| 亚型 | IgG |
| 性状 | Liquid |
| 纯化方法 | affinity purified by Protein A |
| 克隆类型 | Polyclonal |
| 理论分子量 | 178 kDa |
| 浓度 | 1mg/ml |
| 储存液 | 0.01M TBS (pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol. |
| 研究领域 | Epigenetics and Nuclear Signaling > DNA methylation > DNMTs |
| 亚基 | Binds to CSNK1D (By similarity). Homodimer. Interacts with HDAC1 and with PCNA. Forms a complex with DMAP1 and HDAC2, with direct interaction. Forms also a stable complex with E2F1, BB1 and HDAC1. Binds MBD2 and MBD3. Component of complexes containing SUV39H1. Interacts with DNMT3A and DNMT3B. Interacts with the PRC2/EED-EZH2 complex. Interacts with UBC9 and BAZ2A/TIP5. |
| 亚细胞定位 | Nucleus. |
| 组织特异性 | Ubiquitous; highly expressed in fetal tissues, heart, kidney, placenta, peripheral blood mononuclear cells, and expressed at lower levels in spleen, lung, brain, small intestine, colon, liver, and skeletal muscle. Isoform 2 is less expressed than isoform 1. |
| 翻译后修饰 | Sumoylated; sumoylation increases activity.
Acetylation on multiple lysines, mainly by KAT2B/PCAF, regulates cell cycle G(2)/M transition. Deacetylation of Lys-1349 and Lys-1415 by SIRT1 increases methyltransferase activity. Phosphorylation of Ser-154 by CDKs is important for enzymatic activity and protein stability. Phosphorylation of Ser-143 by AKT1 prevents methylation by SETD7 therebye increasing DNMT1 stability. Methylation at Lys-142 by SETD7 promotes DNMT1 proteasomal degradation. |
| 相似性 | Belongs to the C5-methyltransferase family. Contains 2 BAH domains. Contains 1 CXXC-type zinc finger. |
| 功能 | Methylates CpG residues. Preferentially methylates hemimethylated DNA. Associates with DNA replication sites in S phase maintaining the methylation pattern in the newly synthesized strand, that is essential for epigenetic inheritance. Associates with chromatin during G2 and M phases to maintain DNA methylation independently of replication. It is responsible for maintaining methylation patterns established in development. DNA methylation is coordinated with methylation of histones. Mediates transcriptional repression by direct binding to HDAC2. In association with DNMT3B and via the recruitment of CTCFL/BORIS, involved in activation of BAG1 gene expression by modulating dimethylation of promoter histone H3 at H3K4 and H3K9. |
| 保存条件 | Shipped at 4℃. Store at -20℃ for one year. Avoid repeated freeze/thaw cycles. |
| 注意事项 | This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications. |
| 背景资料 | DNA (cytosine-5-)-methyltransferase 1 has a role in the establishment and regulation of tissue-specific patterns of methylated cytosine residues. Aberrant methylation patterns are associated with certain human tumors and developmental abnormalities. Two transcript variants encoding different isoforms have been found for this gene. [provided by RefSeq, Aug 2008]. |
| 应用 | 推荐稀释比例 |
| {IHC-P} | {1:100-500} |
| {IHC-F} | {1:100-500} |
| {IF} | {1:100-500} |
| {Flow-Cyt} | {1ug/Test} |
Paraformaldehyde-fixed, paraffin embedded (human tonsil); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (phospho-Dnmt1 (Ser154)) Polyclonal Antibody, Unconjugated (bs-5294R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (Human kidney); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (phospho-Dnmt1 (Ser154)) Polyclonal Antibody, Unconjugated (bs-5294R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
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文献和实验该产品被引用文献
[IF={{ 2.3 }}] {Tan, Xiaoying, et al. "High inorganic phosphate causes DNMT1 phosphorylation and subsequent fibrotic fibroblast activation." Biochemical and Biophysical Research Communications (2016).} {IP} {="Human"}
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：使用 Anti-phospho-Akt (Ser473) Rabbit mAb 对石蜡包埋的人乳腺癌组织进行免疫组织化学分析。（图 A）使用免疫组化试剂盒M&R HRP/DAB Detection IHC Kit，抗体 1:100 稀释；(图 B) 采用普通免疫组化试剂盒，抗体 1:25 稀释。 图 6 免疫组化实验检测 Erk1/2 表达 注：使用 Anti-Erk1/2 Mouse mAb与p44/42 MAPK （Erk1/2）Rabbit mAb 对正常小鼠心脏组织进行免疫
at 10, 25, and 50 μM). At 24 h post-treatment, celllysates were obtained; SDS-PAGE and Western blotting were performed as described in Section 3, Methods. AR-A014418dose-dependent decreases of phospho-β-catenin (Ser33/37) and phospho-glycogen synthase
(Ser473)(193H12)Rabbit mAb，1:300,稀释，二抗是驴抗兔，1：2000，我是实验室的新手，刚呆俩月，受 此挫折，一头雾水，请过来人或者有经验的同学帮忙解答怎样解决，非常感谢！ 碧峤 pAkt有两个位点，Ser473和Thr308，其中Ser473相对容易显色。如果显色失败，可能的原因:1 上样量过少，我们一般都是30μg；2 一抗孵育时间过短，我们一般24h~48h；3 一抗是否工作有待排除 米宝
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phospho-Dnmt1 (Ser154) Rabbit pAb(bs-5294R)-50ul/100ul/200ul
￥1180 - 2800
