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#12656 Smad 1/5/9 Antibody Sampler Kit

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2019年3月20日11時40分 現在
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キット内容 容量 用途 種交差性 ホモロジー† 検出分子量 アイソタイプ
Phospho-Smad1 (Ser463/465)/ Smad5 (Ser463/465)/ Smad9 (Ser465/467) (D5B10) Rabbit mAb #13820 20 µl WB, IP, IF-IC, F H, M, R Mk 60 Rabbit IgG
Phospho-Smad1/5 (Ser463/465) (41D10) Rabbit mAb #9516 20 µl WB, IF-IC, F H, M, R 60 Rabbit
Phospho-Smad1 (Ser206) (D40B7) Rabbit mAb #5753 20 µl WB, IP H M, R, Mk 60 Rabbit IgG
Smad1 (D59D7) XP® Rabbit mAb #6944 20 µl WB, IP, ChIP H, M, Mk X, B 60 Rabbit IgG
Smad4 (D3M6U) Rabbit mAb #38454 20 µl WB, IP, ChIP, ChIP-seq H, M, R, Mk Dg, GP, Hr 70 Rabbit IgG
Smad5 (D4G2) Rabbit mAb #12534 20 µl WB, IP, ChIP H, M, R, Mk 60 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody #7074 100 µl WB Goat

†Species predicted to react based on 100% sequence homology.

Applications Key: W=Western Blotting, IP=Immunoprecipitation, IF-IC=Immunofluorescence (Immunocytochemistry), F=Flow Cytometry, ChIP=Chromatin IP, ChIP-seq=Chromatin IP-seq
Reactivity Key: H=Human, M=Mouse, R=Rat, Mk=Monkey

社内データ

Western Blotting

Western Blotting

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 Blotting

Western Blotting

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.

Western Blotting

Western Blotting

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 Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Smad1 (D59D7) XP® Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Smad5 (D4G2) Rabbit mAb.

Western Blotting

Western Blotting

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 (Ser463/465)/ Smad5 (Ser463/465)/ Smad9 (Ser465/467) (D5B10) Rabbit mAb (upper) and Smad1 (D59D7) XP® Rabbit mAb #6944 (lower).


Western Blotting

Western Blotting

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.

Chromatin IP-seq

Chromatin IP-seq

Chromatin immunoprecipitations were performed with cross-linked chromatin from HaCaT cells treated with TGF-β1 #8915 (7 ng/mL, 1 hr) and Smad4 (D3M6U) Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. The figure shows binding across ID1, a known target gene of Smad4 (see additional figure containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product data sheet.

Flow Cytometry

Flow Cytometry

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 Blotting

Western Blotting

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).

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from MCF7 cells treated with Human BMP2 #4697 (50 ng/ml) for one hour and either Smad1 (D59D7) XP® Rabbit mAb or 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.

IP

IP

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.


Flow Cytometry

Flow Cytometry

Flow cytometric analysis of HT-1080 cells, untreated (blue) or treated with Human BMP2 #4697 (green), using Phospho-Smad1 (Ser463/465)/ Smad5 (Ser463/465)/ Smad9 (Ser465/467) (D5B10) Rabbit mAb. Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.

IP

IP

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 IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from HaCaT cells treated with TGF-β1 #8915 (7 ng/mL, 1 hr) and either Smad4 (D3M6U) Rabbit mAb or 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).


Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from MCF7 cells treated with Human BMP2 #4697 (50 ng/ml, 1 hr) and either Smad5 (D4G2) Rabbit mAb or 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.

IF-IC

IF-IC

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 (Ser463/465)/ Smad5 (Ser463/465)/ Smad9 (Ser465/467) (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.

IF-IC

IF-IC

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).


IF-IC

IF-IC

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).

バックグラウンド

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) .

使用文献
 
関連製品
12052   Smad2/3 Control Cell Extracts
4680   BMP4 (6B7) Mouse mAb
4693   BMP7 Antibody
4697   Human BMP-2
6223   SignalSilence® Smad1 siRNA I

Illumina is a registered trademark of Illumina, Inc.
Tween is a registered trademark of ICI Americas, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
SimpleChIP is a registered trademark of Cell Signaling Technology, Inc.
LumiGLO is a registered trademark of Kirkegaard & Perry Laboratories.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.

本製品は試験研究用です。

Smad 1/5/9 Antibody Sampler Kit

Immune Cell Signaling Pathways

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