Cell Signaling Technology

Product Pathways - TGF-beta/Smad Signaling

Phospho-Smad1 (Ser463/465)/ Smad5 (Ser463/465)/ Smad9 (Ser465/467) (D5B10) Rabbit mAb (ChIP formulated) #11971

smad   smad1   smad158   smad159   smad5   smad8   smad9  

No. Size Price
11971S 100 µl ( 10 immunoprecipitations ) ¥3,900.00 现货查询 购买询价
11971 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
ChIP 1:100 Human, Endogenous Rabbit IgG

Species cross-reactivity is determined by western blot.

Applications Key: ChIP=Chromatin IP,

Homology

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

Specificity / Sensitivity

Phospho-Smad1/5 (Ser463/465) (D5B10) Rabbit mAb (ChIP formulated) recognizes endogenous levels of Smad1 and Smad5 protein when phosphorylated at Ser463/465.

Phospho-Smad1/5 (Ser463/465) (D5B10) Rabbit mAb (ChIP 级)可以识别内源性总的 Ser463/465被磷酸化的Smad1和Smad5蛋白。

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肽段免疫动物产生,该肽段与人Smad1和Smad5蛋白Ser463/465邻近氨基酸残基序列一致。

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 MCF7 cells treated with Human BMP2 #4697 (50 ng/ml) for 1 hr and either 5 μl of Phospho-Smad1/5 (Ser463/465) (D5B10) Rabbit mAb or 2 μl of Normal Rabbit IgG #2729 using SimpleChIp® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human ID1 Promoter Primers #5139, human SMAD6 promoter primers, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

使用SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003将Human BMP2 #4697 (50 ng/ml)处理1小时的4 x 106 MCF7细胞与5μl的Phospho-Smad1/5 (Ser463/465) (D5B10) Rabbit mAb或2μl的Normal Rabbit IgG #2729交联然后进行染色质免疫共沉淀实验。富集的DNA使用SimpleChIP® Human ID1 Promoter 引物 #5139 human SMAD6 promoter 引物,和SimpleChIP® Human α Satellite Repeat 引物 #4486以real-time PCR的方式定量。各样品沉淀得到的DNA量相对于input染色质总量进行相对定量,Input中染色质量设定为1。

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.

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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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