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Primary Antibodies  >  Transcription Factor Antibodies
 

SMAD3 phospho S423/phospho S425 Antibody

Rabbit Polyclonal
Code

600-401-919
Size

100 µg
Price

$309.00
Availability

Ships next business day.
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Rockland's affinity purified anti-Smad3 pS423 pS425 antibody was used at 2.5 ug/ml to detect signal in a variety of tissues including multi-human, multi-brain and multi-cancer slides. This image shows strong nuclear staining in the majority of epidermal keratinocytes at 40X.  Tissue was formalin-fixed and paraffin embedded.  The image shows localization of the antibody as the precipitated red signal, with a hematoxylin purple nuclear counterstain.  Personal Communi-cation, Tina Roush, LifeSpanBiosciences, Seattle, WA.
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Synonyms: hMAD 3 antibody, hSMAD3 antibody, MADH3 antibody, MGC60396 antibody, Mothers against decapentaplegic homolog 3 antibody, Mothers against DPP homolog 3 antibody

SMAD3 phospho S423/phospho S425 Antibody Properties

Description
Anti-SMAD3 pS423 pS425 (RABBIT) Antibody - 600-401-919
Host
Rabbit
Target Species
Human
Known Cross Reactivity
human, Xenopus laevis, Xenopus tropicalis, zebrafish, rat, mouse, swine, bovine, chicken
Clonality
Polyclonal
Application
ELISA : 1:75,000 - 1:300,000
FlowCytometry : User Optimized
Western Blot : 1:2,000 - 1:20,000
Immunochemistry: 1:500
Other Dilution: User Optimized
Physical State
Liquid (sterile filtered)
Shipping condition
Dry Ice
Format
IgG
Label
Concentration
1.0 mg/mL by UV absorbance at 280 nm
Buffer
0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
Preservative
0.01% (w/v) Sodium Azide
Stabilizer
None
GeneName
SMAD3
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Lot#  

SMAD3 phospho S423/phospho S425 Antibody Description

Background
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.
Immunogen
This affinity purified antibody was prepared from whole rabbit serum produced by repeated immunizations with a synthetic peptide corresponding to amino acids 417-425 of human SMAD3 protein.
Immunogen Type
Peptide
Post Translational Modification
Phosphorylation
Storage Condition
Store vial at -20° C prior to opening. Aliquot contents and freeze at -20° C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4° C as an undiluted liquid. Dilute only prior to immediate use.
Application Note
This affinity purified antibody has been tested for use in ELISA, immunohistochemistry, and 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.
Purity/Specifity
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.
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Product Type
Primary Antibodies;
Applications
ELISA, WB
Reacts With
human, mouse, rat
Catalog Number
600-401-281

Product Type
Primary Antibodies;
Applications
ELISA, WB, IHC
Reacts With
human, Xenopus laevis, Xenopus tropicalis, zebrafish, rat, mouse, swine, bovine, chicken
Catalog Number
600-401-920

Product Type
Primary Antibodies;
Applications
ELISA, WB
Reacts With
human, mouse, rat
Catalog Number
600-401-973

Product Type
Primary Antibodies;
Applications
ELISA, WB
Reacts With
human, mouse, Xenopus
Catalog Number
600-401-A41

Product Label

 

Description
Unconjugated
Color
Conjugation Reference
Molecular Weight
Excitation Wavelength
Conjugation Chemistry