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PS1023 PhosphoDetect™ Anti-Smad3 (pSer423/425) Rabbit mAb (C25A9)

PS1023
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Overview

Replacement Information

Key Specifications Table

Species ReactivityHostAntibody Type
H Rb Monoclonal Antibody

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PS1023-50UL
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      Description
      OverviewRecognizes the ~52 kDa Smad3 protein phosphorylated at Ser423/425 in HT-1080 cells treated with TGF-β1.
      Catalogue NumberPS1023
      Brand Family Calbiochem®
      Application Data
      Detection of human Smad3, phosphorylated at Ser423/425, by immunoblotting. Samples: whole cell lysate from HT-1080 cells, left untreated or treated with TGF-β1, TGFR inhibitor (SB-431542), or BMP-2, as indicated by the (+) and (-). Primary antibody: PhosphoDetect™ Anti-Smad3 (pSer423/425) Rabbit mAb (C25A9) (Cat. No. PS1023) (1:1000) (top panel) and Anti-Smad3 (total) (bottom panel). Detection: chemiluminescence.
      References
      ReferencesAttisano, L., et al. 2002. Science 296, 1646.
      Moustakas, A., et al. 2001. J. Cell Sci. 114, 4359.
      Wrana, J. 2000. Science 23, 1.
      Product Information
      FormLiquid
      FormulationIn 150 mM NaCl, 10 mM HEPES, 100 µg/ml BSA, 50% glycerol pH 7.5.
      Preservative≤0.1% sodium azide
      Quality LevelMQ100
      Applications
      Key Applications Immunoblotting (Western Blotting)
      Immunoprecipitation
      Application NotesImmunoblotting (1:1000)
      Immunoprecipitation (1:50)
      Application CommentsDoes not cross-react with other Smad proteins. For immunoblotting incubate the membrane with antibody diluted with TBS containing 5% BSA, 0.1% Tween®-20 detergent overnight at 4°C with gentle shaking. Please note that this is a rabbit monoclonal antibody and requires the use of an anti-rabbit secondary antibody. The immunogen sequence is 100% conserved in mouse, rat, monkey, bovine, xenopus, and zebrafish Smad3, but cross-reactivity has not been tested. Antibody should be titrated for optimal results in individual systems.
      Biological Information
      Immunogena synthetic phosphopeptide corresponding to amino acids surrounding the Ser 423/425 phosphorylation sites of Smad3, coupled to KLH.
      CloneC25A9
      HostRabbit
      IsotypeIgG
      Species Reactivity
      • Human
      Antibody TypeMonoclonal Antibody
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Storage and Shipping Information
      Ship Code Blue Ice Only
      Toxicity Standard Handling
      Storage -20°C
      Do not freeze Ok to freeze
      Special InstructionsDo not aliquot.
      Packaging Information
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      PhosphoDetect™ Anti-Smad3 (pSer423/425) Rabbit mAb (C25A9) SDS

      Title

      Safety Data Sheet (SDS) 

      PhosphoDetect™ Anti-Smad3 (pSer423/425) Rabbit mAb (C25A9) Certificates of Analysis

      TitleLot Number
      PS1023

      References

      Reference overview
      Attisano, L., et al. 2002. Science 296, 1646.
      Moustakas, A., et al. 2001. J. Cell Sci. 114, 4359.
      Wrana, J. 2000. Science 23, 1.
      Data Sheet

      Note that this data sheet is not lot-specific and is representative of the current specifications for this product. Please consult the vial label and the certificate of analysis for information on specific lots. Also note that shipping conditions may differ from storage conditions.

      Revision20-April-2009 JSW
      ApplicationImmunoblotting (1:1000)
      Immunoprecipitation (1:50)
      Application Data
      Detection of human Smad3, phosphorylated at Ser423/425, by immunoblotting. Samples: whole cell lysate from HT-1080 cells, left untreated or treated with TGF-β1, TGFR inhibitor (SB-431542), or BMP-2, as indicated by the (+) and (-). Primary antibody: PhosphoDetect™ Anti-Smad3 (pSer423/425) Rabbit mAb (C25A9) (Cat. No. PS1023) (1:1000) (top panel) and Anti-Smad3 (total) (bottom panel). Detection: chemiluminescence.
      DescriptionPurified rabbit monoclonal antibody generated by immunizing rabbits with the specified immunogen and fusing splenocytes with an immortal cell line. Recognizes the ~52 kDa Smad3 protein phosphorylated at Ser423/425.
      BackgroundMembers of the Smad family of signal transduction molecules are components of a critical intracellular pathway that transmits TGF-β signals from the cell surface to the nucleus. Three distinct classes of Smads have been defined: the receptor-regulated Smads (R-Smads), which include Smad1, 2, 3, 5 and 8, the common-mediator Smad (co-Smad), Smad4, and the antagonistic or inhibitory Smads (I-Smads), Smad6 and 7. Activated type I receptors associate with specific R-Smads and phosphorylate them on a conserved carboxy-terminal SSXS motif. The phosphorylated R-Smad dissociates from the receptor and forms a heteromeric complex with the co-Smad (Smad4), allowing translocation of the complex to the nucleus. Once in the nucleus, Smads can target a variety of DNA binding proteins to regulate transcriptional responses. Following stimulation by TGF-β, Smad2 and Smad3 become phosphorylated at carboxyl terminal serine residues (Ser465/467 on Smad2; Ser423/425 on Smad3) by TGF-β Receptor I. Phosphorylated Smad 2/3 can complex with Smad4 and translocate to the nucleus to regulate gene expression.
      HostRabbit
      Immunogena synthetic phosphopeptide corresponding to amino acids surrounding the Ser 423/425 phosphorylation sites of Smad3, coupled to KLH.
      CloneC25A9
      IsotypeIgG
      Specieshuman
      FormLiquid
      FormulationIn 150 mM NaCl, 10 mM HEPES, 100 µg/ml BSA, 50% glycerol pH 7.5.
      Preservative≤0.1% sodium azide
      CommentsDoes not cross-react with other Smad proteins. For immunoblotting incubate the membrane with antibody diluted with TBS containing 5% BSA, 0.1% Tween®-20 detergent overnight at 4°C with gentle shaking. Please note that this is a rabbit monoclonal antibody and requires the use of an anti-rabbit secondary antibody. The immunogen sequence is 100% conserved in mouse, rat, monkey, bovine, xenopus, and zebrafish Smad3, but cross-reactivity has not been tested. Antibody should be titrated for optimal results in individual systems.
      Storage -20°C
      Do Not Freeze Ok to freeze
      Special InstructionsDo not aliquot.
      Toxicity Standard Handling
      ReferencesAttisano, L., et al. 2002. Science 296, 1646.
      Moustakas, A., et al. 2001. J. Cell Sci. 114, 4359.
      Wrana, J. 2000. Science 23, 1.