Cell Signaling Technology

Product Pathways - TGF-beta/Smad Signaling

Phospho-Smad1 (Ser463/465)/ Smad5 (Ser463/465)/ Smad9 (Ser465/467) (D5B10) Rabbit mAb (PE Conjugate)  #12428

Smad   smad1   smad158   smad159   smad5   smad8   smad9  

Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
F 1:50 Human,Mouse,Rat, Endogenous Rabbit IgG

Species cross-reactivity is determined by western blot.

Applications Key: F=Flow Cytometry,

Homology

Species predicted to react based on 100% sequence homology: Monkey,

Specificity / Sensitivity

Phospho-Smad1/5 (Ser463/465) (D5B10) Rabbit mAb (PE Conjugate) recognizes endogenous levels of Smad1 protein when phosphorylated at Ser465 only, at both Ser463 and Ser465, or at Ser462, Ser463 and Ser465. This antibody also recognizes the endogenous levels of Smad5 protein when phosphorylated at Ser463 only, Ser465 only, at both Ser463 and Ser465, at both Ser462 and Ser465, or at Ser462, Ser463 and Ser465.

Phospho-Smad1/5 (Ser463/465) (D5B10) Rabbit mAb 兔单抗(PE 偶联)可以识别内源性的磷酸化Smad1蛋白,但是需要Ser465被单独磷酸化,或Ser463和Ser465,被双重磷酸化,或Ser462, Ser463 和 Ser465同时磷酸化。抗体也能识别内源性的磷酸化Smad5蛋白,但是需要Ser463被单独磷酸化,或Ser465被单独磷酸化,或Ser463 和 Ser465同时磷酸化,或Ser462 和 Ser465同时磷酸化,或Ser462, Ser463 和 Ser465被同时磷酸化。

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser463/465 of human Smad1 and Smad5 protein.

此单克隆抗体由合成磷酸肽段免疫动物产生,该重组蛋白与人Smad1和Smad5蛋白Ser463/465邻近氨基酸残基序列一致。

Description

This Cell Signaling Technology antibody is conjugated to phycoerythrin (PE) and tested in-house for direct flow cytometry analysis in human cells. The antibody is expected to exhibit the same species cross-reactivity as the unconjugated Phospho-Smad1/5 (Ser463/465) (D5B10) Rabbit mAb #11971.

此Cell Signaling Technology公司抗体偶联藻红蛋白(PE)并经人类细胞直接流式细胞分析内部测试。抗体预期和未偶联的Phospho-Smad1/5 (Ser463/465) (D5B10) Rabbit mAb兔单抗 #11971显示同样的物种交叉反应

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of HT-1080 cells, untreated (blue) or treated with Human BMP2 #4697 (50 ng/ml, 30 min; green), using Phospho-Smad1/5 (Ser463/465) (D5B10) Rabbit mAb (PE Conjugate).

未处理(蓝色)或经过Human BMP2 #4697 (50 ng/ml, 30 min; 绿色)处理的HT-1080细胞,使用Phospho-Smad1/5 (Ser463/465) (D5B10) Rabbit mAb兔单抗 (PE 偶联)进行流式细胞分析。

Background

Bone morphogenetic proteins (BMPs) constitute a large family of signaling molecules that regulate a wide range of critical processes including morphogenesis, cell-fate determination, proliferation, differentiation, and apoptosis (1,2). BMP receptors are members of the TGF-β family of Ser/Thr kinase receptors. Ligand binding induces multimerization, autophosphorylation, and activation of these receptors (3-5). They subsequently phosphorylate Smad1 at Ser463 and Ser465 in the carboxy-terminal motif SSXS, as well as Smad5 and Smad8 at their corresponding sites. These phosphorylated Smads dimerize with the coactivating Smad4 and translocate to the nucleus, where they stimulate transcription of target genes (5).

MAP kinases and CDKs 8 and 9 phosphorylate residues in the linker region of Smad1, including Ser206. The phosphorylation of Ser206 recruits Smurf1 to the linker region and leads to the degradation of Smad1 (6). Phosphorylation of this site also promotes Smad1 transcriptional action by recruiting YAP to the linker region (7).

骨形成蛋白由一大类信号分子组成,这些分子调控包括形态发生、细胞命运定向、增殖、分化以及凋亡在内的广泛重要过程[1,2]。BMP 受体为TGF-β家族丝氨酸/苏氨酸受体成员。配基的结合诱导多聚化、自磷酸化和相应受体的激活[3-5]。它们随后磷酸化Smad1蛋白的C-末端SSXS模体中的463位丝氨酸和465位丝氨酸,且磷酸化Smad5和Smad8中相应的位置。这些磷酸化的Smad与共激活的Smad4形成二聚体,转移到核内激活靶基因的转录[5]。促分裂原活化蛋白激酶MAPK和周期蛋白依赖性蛋白激酶8和9在Smad1的连接区域磷酸化相应残基包括丝氨酸206。进而募集Smurf1到接头区域导致Smad1的降解[6]。这一位点的磷酸化也招募YAP到接头区域促进Smad1的转录激活[7]。

  1. Hogan, B.L. (1996) Genes Dev 10, 1580-94.
  2. Hoodless, P.A. et al. (1996) Cell 85, 489-500.
  3. Klemm, J.D. et al. (1998) Annu Rev Immunol 16, 569-92.
  4. Kretzschmar, M. et al. (1997) Genes Dev 11, 984-95.
  5. Whitman, M. (1998) Genes Dev 12, 2445-62.
  6. Sapkota, G. et al. (2007) Mol Cell 25, 441-54.
  7. Alarcón, C. et al. (2009) Cell 139, 757-69.

Application References

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For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.

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