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- 详细信息
- 文献和实验
- 技术资料
- 亚型:
IgG
- 保存条件:
Store at -20°C. Avoid exposure to light. Stable for one year after shipment.
- 克隆性:
Recombinant
- 标记物:
CoraLite® Plus 488 Fluorescent Dye
- 适应物种:
human
- 宿主:
Rabbit
- 应用范围:
FC (Intra)
- 靶点:
Phospho-SMAD2 (Ser465/467)/SMAD3 (Ser423/425)
- 抗体英文名:
CoraLite® Plus 488-conjugated Phospho-SMAD2 (Ser465/467)/SMAD3 (Ser423/425) Recombinant monoclonal antibody
- 抗体名:
CoraLite® Plus 488-conjugated Phospho-SMAD2 (Ser465/467)/SMAD3 (Ser423/425) Recombinant monoclonal antibody
- 规格:
100ul
经过测试的应用
| Positive FC (Intra) detected in | Calyculin A treated HEK-293 cells |
推荐稀释比
| 应用 | 推荐稀释比 |
|---|---|
| Flow Cytometry (FC) (INTRA) | FC (INTRA) : 0.25 ug per 10^6 cells in a 100 µl suspension |
| It is recommended that this reagent should be titrated in each testing system to obtain optimal results. | |
| Sample-dependent, Check data in validation data gallery. | |
产品信息
CL488-80427-2 targets Phospho-SMAD2 (Ser465/467)/SMAD3 (Ser423/425) in FC (Intra) applications and shows reactivity with human samples.
| 经测试应用 | FC (Intra) |
| 经测试反应性 | human |
| 免疫原 |
fusion protein |
| 宿主/亚型 | Rabbit / IgG |
| 抗体类别 | Recombinant |
| 产品类型 | Antibody |
| 全称 | SMAD family member 2 |
| 别名 | SMAD2, JV18 1, JV18, hSMAD2, hMAD-2 |
| 计算分子量 | 467 aa, 52 kDa |
| 观测分子量 | 60 kDa |
| GenBank蛋白编号 | BC014840 |
| 基因名称 | SMAD2 |
| Gene ID (NCBI) | 4087 |
| 偶联类型 | CoraLite® Plus 488 Fluorescent Dye |
| 最大激发/发射波长 | 493 nm / 522 nm |
| 形式 | Liquid |
| 纯化方式 | Protein A purification |
| UNIPROT ID | Q15796 |
| 储存缓冲液 | PBS with 50% glycerol, 0.05% Proclin300, 0.5% BSA, pH 7.3. |
| 储存条件 | Store at -20°C. Avoid exposure to light. Stable for one year after shipment. Aliquoting is unnecessary for -20oC storage. |
背景介绍
SMAD2, also named as MADH2 and MADR2, belongs to the dwarfin/SMAD family, contains 1 MH1 (MAD homology 1) domain and 1 MH2 (MAD homology 2) domain. SMAD2 is a receptor-regulated SMAD(R-SMAD) that is an intracellular signal transducer and transcriptional modulator activated by TGF-beta and activin type 1 receptor kinases. This protein may act as a tumor suppressor in colorectal carcinoma. It is phosphorylated on one or several of Thr-220, Ser-245, Ser-250, and Ser-255. In response to TGF-beta, It is phosphorylated on Ser-465/467 by TGF-beta and activin type 1 receptor kinases, and then able to interact with SMURF2, recruiting other proteins, such as SNON, for degradation. In response to decorin, the naturally occurring inhibitor of TGF-beta signaling, it is phosphorylated on Ser-240 by CaMK2. It is phosphorylated by MAPK3 upon EGF stimulation; which increases transcriptional activity and stability, and is blocked by calmodulin. In response to TGF-beta, it is ubiquitinated by NEDD4L, which promotes its degradation. In response to TGF-beta signaling, it is acetylated on Lys-19 by coactivators, which increases transcriptional activity. The molecular weight of unphosphorylated forms of Smad2 is 52 kDa and phosphorylated forms of Smad2 is 58 kDa. (PMID: 9006934)
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文献和实验下调不能说明活性降低,因为决定GSK3-beta的活性的还与其磷酸化的比例和磷酸化的位点有关。所以你除了要做GSK3-beta总的蛋白表达之外,还要做磷酸化的GSK3-beta的蛋白表达。 (2)应该活性形式和非活性形式都做。一般来讲,GSK3-beta在Ser9位点磷酸化之后活性收到抑制,而在216位点磷酸化之后,其活性收到加强。因此建议将GSK3-beta的两个磷酸化位点都做了,另外还要同时检测GSK3-beta的总蛋白表达,这样才能全面的说明问题。
TBS 和 5% NGS 中封闭样品。市售的含有酪蛋白的封闭溶液与磷酸化的一抗结合后易减弱信号;因此,我们建议不使用含有酪蛋白的封闭剂进行磷酸化特异性抗体的检测。以上针对封闭步骤所提建议均是在以 SignalStain. Boost IHC Detection Reagent 作为检测试剂的情况下提出的。如果选择使用其他检测试剂,我们则建议使用与二抗来源相同的血清进行封闭。抗体:Phospho–Histone H2A.X (Ser139) (20E3) Rabbit mAb #9718样本:石蜡
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
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