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SMAD3 phospho S423/phospho S425 Antibody

Rabbit Polyclonal
JCYIA Product
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  • Immunohistochemistry - SMAD3pS423pS425
  • Western Blot - Anti-Smad3 pS423 pS425 Antibody
25 µL
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SMAD3 phospho S423/phospho S425 Antibody Properties

Anti-SMAD3 pS423 pS425 (RABBIT) Antibody - 600-401-919S
Target Species
Known Cross Reactivity
human, Xenopus laevis, Xenopus tropicalis, zebrafish, rat, mouse, swine, bovine, chicken
ELISA : 1:75,000 - 1:300,000
Flow Cytometry : User Optimized
Western Blot : 1:2,000 - 1:20,000
Immunohistochemistry: 1:500
Other Dilution: User Optimized
Physical State
Liquid (sterile filtered)
Shipping Condition
Dry Ice
1.0 mg/mL by UV absorbance at 280 nm
0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
0.01% (w/v) Sodium Azide
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SMAD3 phospho S423/phospho S425 Antibody Description

This antibody is designed, produced, and validated as part of a collaboration between Rockland and the National Cancer Institute (NCI) and is suitable for Cancer, Immunology and Nuclear Signaling research. Smad3 (also known as Mothers against decapentaplegic homolog 3 Mothers against DPP homolog 3, Mad3, hMAD-3, JV15-2 or hSMAD3) is a transcriptional modulator activated by TGF-beta (transforming growth factor) and activin type 1 receptor kinase. These activators exert diverse effects on a wide array of cellular processes. The Smad proteins mediate much of the signaling responses induced by the TGF-b superfamily. Briefly, activated type I receptor phosphorylates receptor-activated Smads (R-Smads) at their c-terminal two extreme serines in the SSXS motif, e.g. Smad2 and Smad3 proteins in the TGF-b pathway, or Smad1, Smad5 or Smad8 in the BMP pathway. Then the phosphorylated R-Smad translocated into nucleus, where they regulate transcription of target genes. Based on microarray and animal model experiments, Smad3 accounts for at least 80% of all TGF-b-mediated response.
rabbit anti-SMAD3 pS423pS425 antibody, SMAD-3, SMAD 3, mothers against decapentaplegic homolog 3 antibody, MAD homolog 3, Mothers against DPP homolog 3, SMAD family member 3, MADH3, MADH 3, JV15-2
Anti-SMAD3 pS423pS425 antibody was prepared from whole rabbit serum produced by repeated immunizations with a dual phosphorylated synthetic peptide corresponding to a c-terminal region with Serine 423 and Serine 425 of human SMAD3 protein.
Immunogen Type
Post Translational Modification
Storage Condition
Store vial at -20° C or below prior to opening. This vial contains a relatively low volume of reagent (25 µL). To minimize loss of volume dilute 1:10 by adding 225 µL of the buffer stated above directly to the vial. Recap, mix thoroughly and briefly centrifuge to collect the volume at the bottom of the vial. Use this intermediate dilution when calculating final dilutions as recommended below. Store the vial at -20°C or below after dilution. Avoid cycles of freezing and thawing.
Application Note
This affinity purified antibody has been tested for use in ELISA, immunohistochemistry and by western blot. Specific conditions for reactivity should be optimized by the end user. Expect a band approximately 48 kDa in size corresponding to phosphorylated Smad3 protein by western blotting in the appropriate stimulated tissue or cell lysate or extract. Less than 0.2% reactivity is observed against the non-phosphorylated form of the immunizing peptide. This antibody is phospho specific for dual phosphorylated pS423 and pS425 of Smad3. Stimulation with 2 ng/ml TGF-beta for 1 hour is suggested.
This affinity-purified antibody is directed against the phosphorylated form of human Smad3 protein at the pS423 and pS425 residues. The product was affinity purified from monospecific antiserum by immunoaffinity purification. Antiserum was first purified against the phosphorylated form of the immunizing peptide. The resultant affinity purified antibody was then cross adsorbed against the non-phosphorylated form of the immunizing peptide. Reactivity occurs against human Smad3 pS423 and pS425 protein and the antibody is specific for the phosphorylated form of the protein. Reactivity with non-phosphorylated human Smad3 is minimal by ELISA and western blot. Expect reactivity against phosphorylated Smad1 and Smad5. Negligible reactivity is seen against other phosphorylated Smad family members. A BLAST analysis was used to suggest cross reactivity with Smad3 from human, Xenopus laevis, Xenopus tropicalis, zebrafish, rat, mouse, swine, bovine and chicken based on 100% sequence homology with the immunogen. Reactivity against homologues from other sources is not known.
Disclaimer Note-General
This product is for research use only and is not intended for therapeutic or diagnostic applications. Please contact a technical service representative for more information. All products of animal origin manufactured by Rockland Immunochemicals are derived from starting materials of North American origin. Collection was performed in United States Department of Agriculture (USDA) inspected facilities and all materials have been inspected and certified to be free of disease and suitable for exportation. All properties listed are typical characteristics and are not specifications. All suggestions and data are offered in good faith but without guarantee as conditions and methods of use of our products are beyond our control. All claims must be made within 30 days following the date of delivery. The prospective user must determine the suitability of our materials before adopting them on a commercial scale. Suggested uses of our products are not recommendations to use our products in violation of any patent or as a license under any patent of Rockland Immunochemicals, Inc. If you require a commercial license to use this material and do not have one, then return this material, unopened to: Rockland Inc., P.O. BOX 5199, Limerick, Pennsylvania, USA.
General Reference
Shi Y, Massague J. Mechanisms of TGF-beta signaling from cell membrane to the nucleus. Cell. 2003 Jun 13;113(6):685-700. Review. Derynck R, Zhang YE. Smad-dependent and Smad-independent pathways in TGF-beta family signalling. Nature. 2003 Oct 9;425(6958):577-84. Review. Zhang Y, Feng X, We R, Derynck R. Receptor-associated Mad homologues synergize as effectors of the TGF-beta response. Nature. 1996 Sep 12;383(6596):168-72.
Specific Reference
Yamashita M, Fatyol K, Jin C, Wang X, Liu Z and Zhang YE. (2008) TRAF6 Mediates Smad-Independent Activation of JNK and p38 by TGFß. Molecular Cell 31, 918–924. Herhaus L, Al-Salihi M, Macartney T, Weidlich S, Sapkota GP. (2013) OTUB1 enhances TGFß signalling by inhibiting the ubiquitylation and degradation of active SMAD2/3. Nat Commun. 2013;4:2519. doi: 10.1038/ncomms3519. Tripathi V, Sixt KM, Gao S, Xu X, Huang J, Weigert R, Zhou M, Zhang YE. (2016) Direct Regulation of Alternative Splicing by SMAD3 through PCBP1 Is Essential to the Tumor-Promoting Role of TGF-ß. Mol Cell. 2016 Dec 1;64(5):1010. doi: 10.1016/j.molcel.2016.11.025.
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Product Type
Primary Antibodies;
Reacts With
human, mouse, rat
Catalog Number

Product Type
Primary Antibodies;
Reacts With
human, Xenopus laevis, Xenopus tropicalis, zebrafish, rat, mouse, swine, bovine, chicken
Catalog Number

Product Type
Primary Antibodies;
Reacts With
human, Xenopus laevis, Xenopus tropicalis, zebrafish, rat, mouse, swine, bovine, chicken
Catalog Number

Product Type
Primary Antibodies;
Reacts With
human, mouse, rat
Catalog Number

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Conjugation Reference
Molecular Weight
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