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SMAD2/3 Antibody Sampler Kit
SMAD2/3 Antibody Sampler Kit
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SMAD2/3 Antibody Sampler Kit

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    产品介绍
    产品信息
    抗原名称
    Smad23
    来源纯化
    Phospho-specific monoclonal antibodies are produced by immunizing animals with synthetic phosphopeptides corresponding to residues surrounding Ser465/467 of human SMAD2 or Ser423/425 of human SMAD3 protein. Total SMAD2, SMAD3, and SMAD4 monoclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues near the amino termini of mouse SMAD2 and SMAD3, surrounding His198 of human SMAD2/3, or surrounding Asp165 of human SMAD4 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 SMAD2/3 Antibody Sampler Kit recognizes only its specific target and does not cross-react with other family members. Activation state antibodies detect their intended targets only when phosphorylated at the indicated site. The total SMAD2, SMAD3, and SMAD4 antibodies detect their respective targets at endogenous levels.

    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 Smad1/5/8 for the bone morphogenetic protein (BMP) pathway. Carboxy-terminal phosphorylation of Smad proteins by activated receptors results in their partnering with the common signaling transducer Smad4, and translocation to the nucleus. Activated Smad proteins regulate diverse biological effects by partnering with transcription factors resulting in cell-state specific modulation of transcription (1-4). 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.

    参考图片

    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.

    Western blot analysis of extracts from HT1080 (human), C2C12 (mouse) and B35 (rat) using Smad3 (C67H9) Rabbit mAb.

    Western blot analysis of extracts from HT1080 cells, treated with TGF-β1, TGFR inhibitor SB-431542 or BMP-2, using Phospho-Smad3 (Ser423/425) (C25A9) Rabbit mAb #9520 (upper) or total Smad3 (C67H9) Rabbit mAb #9523 (lower).

    Confocal immunofluorescent analysis of HT1080 cells, untreated (left) or TGFβ-treated (right), using Smad3 (C67H9) Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red).

    Flow cytometric analysis of HT-1080 cells using Smad3 (C67H9) Rabbit mAb #9523 (blue) 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.

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

    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.

    Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HaCaT cells treated with Human TGF-β3 #3706 (7 ng/ml) for 1 h and either 10 μl of Smad3 (C67H9) 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 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.

    Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HaCaT cells treated with Human TGF-β3 #3706 (7 ng/ml) for 1 h and either 5 μl of Phospho-Smad3 (Ser423/425) (C25A9) 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 CDKN1A Intron 1 Primers #4669, SimpleChIP® Human ID1 Promoter Primers #5139, human c-Myc intron 1 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.

    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 NIH/3T3 cells, serum-starved (left) or treated with hTGF-β3 #8425 (right), using Smad2 (D43B4) XP® Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red).

    Western blot analysis of extracts from untreated or TGF-beta treated HeLa and NIH/3T3 cells, using Phospho-Smad2 (Ser465/467) (138D4) Rabbit mAb (upper), or Smad2 Antibody #3102 (lower).

    Western blot analysis of extracts from various cell lines using Smad2 (D43B4) XP® Rabbit mAb.

    Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HaCaT cells treated with Human TGF-β3 #8425 (7 ng/ml) for 1 h and either 10 μl of Smad2 (D43B4) XP® Rabbit mAb #5339 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.

    Flow cytometric analysis of HeLa cells using Smad2 (D43B4) XP® Rabbit mAb (blue) compared to a nonspecific negative control antibody (red).

    Flow cytometric analysis of HeLa cells using Smad2/3 (D7G7) XP® Rabbit mAb (blue) compared to Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (red).

    Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HaCaT cells treated with hTGF-β3 #8425 (7 ng/ml, 1 hr) and either 10 μl of Smad2/3 (D7G7) 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 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 HeLa cells, serum starved (left), treated with hTGF-β3 #8425 (100 ng/ml, 30 min, center), or treated with hTGF-β3 and SB43152 (10 ug/mL, 1 hr, right), using Smad2/3 (D7G7) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red).

    Western blot analysis of extracts from HeLa and ACHN cells using Smad2/3 (D7G7) XP® Rabbit mAb.

    Western blot analysis of extracts from untreated (-) or TGF-β treated (+) HeLa and NIH/3T3 cells using Phospho-Smad2 (Ser465/467) (138D4) Rabbit mAb #3108 (upper) or Smad2 Antibody #3102 (lower).使用Phospho-Smad2 (Ser465/467) (138D4) Rabbit mAb #3108未处理(-)或Smad2抗体#3102（下）对未处理(-)或TGF-β treated (+)处理的HeLa细胞和NIH/3T3细胞提取物进行western blot分析。

    Western blot analysis of extracts from various cell lines using Smad2 (D43B4) XP® Rabbit mAb #5339.使用Smad2 (D43B4) XP®Rabbit mAb#5339对多种细胞提取物进行western blot分析。

    Western blot analysis of extracts from HeLa and ACHN cells using Smad2/3 (D7G7) XP® Rabbit mAb #8685.使用Smad2/3 (D7G7) XP®Rabbit mAb#8685对HeLa和ACHN细胞提取物进行western blot分析。

    Western blot analysis of extracts from HT-1080 (human), C2C12 (mouse) and B35 (rat) cells using Smad3 (C67H9) Rabbit mAb #9523.使用Smad3 (C67H9)Rabbit mAb#9523对HT-1080 (human), C2C12 (mouse)和B35 (rat)提取物进行western blot分析。

    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).

    Western blot analysis of extracts from HT-1080, C2C12, or KNRK cells, untreated (-) or treated with TGF-β (10 ng/ml, 30 min; +), using Phospho-Smad3 (Ser423/425) (C25A9) Rabbit mAb (upper) or total Smad3 (C67H9) Rabbit mAb #9523 (lower).

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    货号：
    12747T
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