| 货号 | 38454S |
| 同种亚型 | Rabbit IgG |
| 反应种属 | Human\Mouse\Rat\Moneky |
| 来源宿主 | Rabbit IgG |
| 应用 | WB, IP , ChIP |
| 目标/特异性 | Smad4 (D3M6U) Rabbit mAb recognizes endogenous levels of total Smad4 protein. |
| 使用方法 | W IP ChIP |
| 供应商 | CST |
| 灵敏度 | Endogenous |
| 背景 | Members of the Smad family of signal transduction molecules are components of a critical intracellular pathway that transmits TGF-β signals from the cell surface into the nucleus. Three distinct classes of Smads have been defined: the recepter-regulated Smads (R-Smads), which include Smad1, 2, 3, 5, 8; the common-mediator Smad (co-Smad), Smad4; and the antagonistic or inhibitory Smads (I-Smads), Smad6 and 7 (1-5). Briefly, activated type I receptors associate with specific R-Smads and phosphorylate them on a conserved SSXS motif at the carboxy-terminus of the proteins. The phosphorylated R-Smad dissociates from the receptor and forms a heteromeric complex with the co-Smad, Smad4, and together the complex moves to the nucleus. Once in the nucleus, Smads can target a variety of DNA binding proteins to regulate transcriptional responses (6-8). |
| 存放说明 | -20C |
| 计算分子量 | 70 |
| 参考文献 | 1 . Heldin, C.H. et al. (1997) Nature 390, 465-71. 2 . Attisano, L. and Wrana, J.L. (1998) Curr Opin Cell Biol 10, 188-94. 3 . Derynck, R. et al. (1998) Cell 95, 737-740. 4 . Massagué, J. (1998) Annu Rev Biochem 67, 753-91. 5 . Whitman, M. (1998) Genes Dev 12, 2445-62. 6 . Wrana, J. (2000) Science 23, 1-9. 7 . Attisano, L. and Wrana, J.L. (2002) Science 296, 1646-7. 8 . Moustakas, A. et al. (2001) J Cell Sci 114, 4359-69. |
Western blot analysis of extracts from various cell lines using Smad4 (D3M6U) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower). HT-29 and COLO 205 are Smad4-null mutant cell lines, confirming specificity of the antibody. | |
Immunoprecipitation of Smad4 protein from HCT 116 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Smad4 (D3M6U) Rabbit mAb. Western blot analysis was performed using Smad4 (D3M6U) Rabbit mAb. | |
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HaCaT cells treated with TGF-β1 #8915 (7 ng/mL, 1 hr) and either 5 µl of Smad4 (D3M6U) 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 JunB 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 (equivalent to one). |
