| 货号 | 12656T |
| 目标/特异性 | Activation state antibodies detect their intended targets only when phosphorylated at the indicated site. The total Smad1, Smad4, and Smad5 antibodies detect their respective targets at endogenous levels. |
| 供应商 | CST |
| 背景 | Transforming growth factor-β (TGF-β) superfamily signaling plays a critical role in the regulation of cell growth, differentiation, and development in a wide range of biological systems. In general, signaling is initiated with ligand-induced oligomerization of serine/ threonine receptor kinases and phosphorylation of the cytoplasmic signaling molecules Smad2 and Smad3 for the TGF-β/activin pathway, or Smad1/5/8 for the bone morphogenetic protein (BMP) pathway. Carboxy-terminal phosphorylation of Smads by activated receptors results in their partnering with the common signaling transducer Smad4, and translocation to the nucleus. Activated Smads regulate diverse biological effects by partnering with transcription factors resulting in cell-state specific modulation of transcription (1-7) . 转化生长因子-β (TGF-β)超家族在调控细胞生长，分化和多种生物系统发育过程中发挥了关键作用。总体来说，信号通路是配体诱导的丝氨酸/苏氨酸受体激酶寡聚化，同时细胞质信号分子Smad2和Smad3磷酸化起始TGF-β/activin信号通路，或是Smad1/5/8磷酸化激活骨形成蛋白（BMP）信号通路。活化受体会磷酸化Smad蛋白的羧基末端会导致它们与通用信号转运子Smad4结合，转位到细胞核内。活化的Smad蛋白通过协同转录因子调控多种生物功能，导致细胞特定状态的转录调控（1-7）。 |
| 存放说明 | -20C |
| 参考文献 | 1 . Horbelt, D. et al. (2012) Int J Biochem Cell Biol 44, 469-74. 2 . Ikushima, H. and Miyazono, K. (2010) Nat Rev Cancer 10, 415-24. 3 . Kitisin, K. et al. (2007) Sci STKE 2007, cm1. 4 . Schmierer, B. and Hill, C.S. (2007) Nat Rev Mol Cell Biol 8, 970-82. 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. |
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO* is added and emits light during enzyme catalyzed decomposition. | |
Western blot analysis of extracts from COS, NIH3T3, PC12, and SK-N-MC cells, using Smad4 Antibody. | |
Flow cytometric analysis of HeLa cells, untreated (blue) or BMP-treated (green), using Phospho-Smad1/5 (Ser463/465) (41D10) Rabbit mAb compared to a nonspecific negative control antibody (red). | |
Western blot analysis of extracts from untreated or BMP-4-treated HeLa or NIH/3T3 cells using Phospho-Smad1/5 (Ser463/465) (41D10) Rabbit mAb. | |
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HaCaT cells treated with Human TGF-β3 #3706 (7ng/ml) for 1 h and either 20 μl of Smad4 Antibody 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 CDKN1A Intron 1 Primers #4669, SimpleChIP® Human ID1 Promoter Primers #5139, 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. | |
Confocal immunofluorescent analysis of HT-1080 cells, BMP-treated (left) and untreated (right), using Phospho-Smad5 (Ser463/Ser465) (41D10) Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). | |
Western blot analysis of extracts of HeLa cells, untreated or UV-treated (60 mJ/cm2 for 2 minutes followed by 1.5 hour recovery), using Phospho-Smad1 (Ser206) (D40B7) Rabbit mAb (upper) and Smad1 Antibody #9743 (lower). | |
Western blot analysis of extracts from HT-1080 cells, untreated or treated with TPA #4174 (200 nM for 30 minutes), using Phospho-Smad1 (Ser206) (D40B7) Rabbit mAb (upper) and Smad1 Antibody #9743 (lower). | |
Western blot analysis of extracts from various cell lines using Smad1 (D59D7) XP® Rabbit mAb. | |
Flow cytometric analysis of HT-1080 cells using Smad1 (D59D7) XP® Rabbit mAb (blue) compared to a nonspecific negative control antibody (red). | |
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 MCF7 cells treated with Human BMP2 #4697 (50 ng/ml) for one hour and either 5 μl of Smad1 (D59D7) XP® 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. | |
Confocal immunofluorescent analysis of HT-1080 cells, untreated (left) or treated with human BMP2 #4697 (right), using Smad1 (D59D7) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). | |
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 MCF7 cells treated with Human BMP2 #4697 (50 ng/ml, 1 hr) and either 5 μl of Smad5 (D4G2) 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. | |
Western blot analysis of extracts from various cell lines using Smad5 (D4G2) Rabbit mAb. | |
Immunoprecipitation of Smad5 from HT-1080 cell extracts using Normal Rabbit IgG #2729 (lane 2) or Smad5 (D4G2) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using Smad5 (D4G2) Rabbit mAb. |
