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SMAD 1/5/9 Antibody Sampler Kit
SMAD 1/5/9 Antibody Sampler Kit
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SMAD 1/5/9 Antibody Sampler Kit

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    产品介绍
    产品信息
    抗原名称
    Smad 159
    来源纯化
    Phospho-specific monoclonal antibodies are produced by immunizing animals with synthetic phosphopeptides corresponding to residues surrounding Ser206 of human SMAD1 protein or Ser463/465 of human SMAD1 and SMAD5 proteins. Total SMAD1, SMAD4, and SMAD5 monoclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues surrounding Ser190 of human SMAD1, Asp165 of human SMAD4, or Pro249 of human SMAD5 protein.
    简单描述
    Antibody Sampler Kit for studying in the research area.
    应用
    目标/特异性

    Specificity/Sensitivity

    Each antibody in the SMAD1/5/9 Antibody Sampler Kit recognizes endogenous levels of its respective target. Activation state antibodies detect their intended targets only when phosphorylated at the indicated site(s).

    背景
    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 Smad1/5/9 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) . 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.

    参考图片

    Western blot analysis of extracts from COS, NIH3T3, PC12, and SK-N-MC cells, using Smad4 Antibody.

    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.

    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).

    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.

    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.

    Western blot analysis of extracts from HeLa cells, untreated (-) or UV-treated (60 mJ/cm2 for 2 min, 1.5 hr recovery; +), using Phospho-Smad1 (Ser206) (D40B7) Rabbit mAb #5753 (upper) and Smad1 Antibody #9743 (lower). 未处理(-)或UV处理(60 mJ/cm2 for 2 min, 1.5 hr recovery; +)的HeLa细胞提取物，使用Phospho-Smad1 (Ser206) (D40B7) Rabbit mAb #5753 (上)和 Smad1 Antibody #9743 (下)进行western blot分析。

    Western blot analysis of extracts from HeLa and NIH/3T3 cells, untreated (-) or BMP-4-treated (+), using Phospho-Smad1/5 (Ser463/465) (41D10) Rabbit mAb #9516. 未处理(-)或BMP4处理(+)的HeLa细胞和NIH/3T3细胞，使用hospho-Smad1/5 (Ser463/465) (41D10) Rabbit mAb #9516进行western blot分析。

    Western blot analysis of extracts from various cell lines using Smad1 (D59D7) XP® Rabbit mAb #6944. 使用Smad1 (D59D7) XP® Rabbit mAb #6944对多种细胞提取物进行western blot分析。

    Western blot analysis of extracts from various cell lines using Smad5 (D4G2) Rabbit mAb #12534. 使用Smad5 (D4G2) Rabbit mAb #12534对多种细胞提取物进行western blot分析。

    Western blot analysis of extracts from NIH/3T3 cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-) or SignalSilence® Smad4 siRNA I (Mouse Specific) (+), using Smad4 Antibody #9515 (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). The Smad4 Antibody confirms silencing of Smad4 expression, while the β-Actin (D6A8) Rabbit mAb is used as a loading control.

    Western blot analysis of extracts from Hep G2 or MEF cells, untreated (-) or treated with Human BMP2 #4697 (50 ng/ml, 30 min; +), using Phospho-Smad1/5 (Ser463/465) (D5B10) Rabbit mAb (upper) and Smad1 (D59D7) XP® Rabbit mAb #6944 (lower).

    Flow cytometric analysis of HT-1080 cells, untreated (blue) or treated with Human BMP2 #4697 (green), using Phospho-Smad1/5 (Ser463/465) (D5B10) Rabbit mAb. Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.

    Confocal immunofluorescent analysis of HT-1080 cells, serum-starved (overnight; left) or serum-starved and treated with Human BMP2 #4697 (50 ng/ml, 30 min; right), using Phospho-Smad1/5 (Ser463/Ser465) (D5B10) Rabbit mAb (green) and β-Actin (13E5) Rabbit mAb (Alexa Fluor® 647 Conjugate) #8584 (red). Blue pseudocolor = Propidium Iodide (PI)/RNase Staining Solution #4087.

    Confocal immunofluorescent analysis of HT1080 cells, serum-starved (left) or serum-starved then treated with hBMP2 (50 ng/ml, 30 min; right) using Phospho-Smad1/5 (Ser463/465) (41D10) Rabbit mAb (green) and Cox IV (4D11-B3-E8) Mouse mAb #11967 (red).

    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).

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    货号：
    12656T
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