Blank control: K562.
Primary Antibody (green line): Rabbit Anti-phospho-MyoD1 (Ser200) antibody (bs-5499R)
Dilution:1μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody : Goat anti-rabbit IgG-FITC
Dilution: 0.5μg /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-20℃.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.
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- 2025年10月24日
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| 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-5499R |
| 英文名称 | phospho-MyoD1 (Ser200) Rabbit pAb |
| 中文名称 | 磷酸化肌原调节蛋白抗体 |
| 英文别名 | MyoD1(phospho Ser200); MyoD1(phospho S200); p-MyoD1(phospho S200); MYOD1(phospho S200); Myogenic differentiation 1; AI503393; bHLHc1; MD1; MGC156574; MYF3; MYOD; MYOD1; PUM; MYOD1_HUMAN. |
| 产品应用 | WB=1:500-2000, Flow-Cyt=1ug/Test Not yet tested in other applications. |
| 交叉反应 | Human, Mouse (Rat, Pig, Cow) |
| 抗体来源 | Rabbit |
| 免疫原 | KLH conjugated Synthesised phosphopeptide derived from human MyoD1 around the phosphorylation site of Ser200 |
| 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 | 34 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 | Developmental Biology > Organogenesis > Skeletal development > Muscle Epigenetics and Nuclear Signaling > Transcription > Domain Families > HLH / Leucine Zipper > HLH |
| 亚基 | Efficient DNA binding requires dimerization with another bHLH protein. Seems to form active heterodimers with ITF-2. Interacts with SUV39H1 and CDK9. Interacts with DDX5 (By similarity). |
| 亚细胞定位 | Nucleus. |
| 翻译后修饰 | Phosphorylated by CDK9. This phosphorylation promotes its function in muscle differentiation. Acetylated by a complex containing EP300 and PCAF. The acetylation is essential to activate target genes. Conversely, its deacetylation by SIRT1 inhibits its function (By similarity). Ubiquitinated on the N-terminus; which is required for proteasomal degradation. |
| 相似性 | Contains 1 bHLH (basic helix-loop-helix) domain. |
| 功能 | Involved in muscle differentiation (myogenic factor). Induces fibroblasts to differentiate into myoblasts. Activates muscle-specific promoters. Interacts with and is inhibited by the twist protein. This interaction probably involves the basic domains of both proteins (By similarity). |
| 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 | This gene encodes a nuclear protein that belongs to the basic helix-loop-helix family of transcription factors and the myogenic factors subfamily. It regulates muscle cell differentiation by inducing cell cycle arrest, a prerequisite for myogenic initiation. The protein is also involved in muscle regeneration. It activates its own transcription which may stabilize commitment to myogenesis. [provided by RefSeq, Jul 2008] |
| 应用 | 推荐稀释比例 |
| {WB} | {1:500-2000} |
| {Flow-Cyt} | {1ug/Test} |
Sample:
Lane 1: Mouse Muscle tissue lysates
Lane 2: Mouse Tongue tissue lysates
Lane 3: Mouse NIH/3T3 cell lysates
Primary: Anti- phospho-MyoD1 (Ser200) (bs-5499R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 34 kDa
Observed band size: 47 kDa
Lane 1: Mouse Muscle tissue lysates
Lane 2: Mouse Tongue tissue lysates
Lane 3: Mouse NIH/3T3 cell lysates
Primary: Anti- phospho-MyoD1 (Ser200) (bs-5499R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 34 kDa
Observed band size: 47 kDa
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文献和实验该产品被引用文献
[IF={{ 4.938 }}] {Du J et al. The regulation of skeletal muscle fiber-type composition by betaine is associated with NFATc1/MyoD.J Mol Med (Berl). 2018 Jun 6.} {WB} {Mouse}
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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-MyoD1 (Ser200) Rabbit pAb(bs-5499R)-50ul/100ul/200ul
￥1180 - 2800
