Smad3 [p Ser423, p Ser425] Antibody

Images

 
Western Blot: Smad3 [p Ser423, p Ser425] Antibody [NBP1-77836] - Shows detection of endogenous Smad3 in stimulated cell lysates. Lysates were prepared from control cells (- lanes), or cells stimulated with 2 ng/ml TGF ...read more
Biological Strategies: Immunocytochemistry/ Immunofluorescence: Smad3 [p Ser423, p Ser425] Antibody [NBP1-77836] - BH4 precursor, sepiapterin, normalizes NO level and ameliorates malignant phenotype of ...read more
Immunohistochemistry: Smad3 [p Ser423, p Ser425] Antibody [NBP1-77836] - 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 ...read more

Product Details

Summary
Reactivity Hu, MuSpecies Glossary
Applications WB, ELISA, Flow, ICC/IF, IHC, IHC-P, Single-Cell Western
Clonality
Polyclonal
Host
Rabbit
Conjugate
Unconjugated
Validated by:
   

Biological Strategies

     

Order Details

Smad3 [p Ser423, p Ser425] Antibody Summary

Immunogen
Smad3 [p Ser423, p Ser425] 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. (Uniprot: P84022)
Modification
p Ser423, p Ser425
Specificity
This affinity-purified antibody is directed against the phosphorylated form of human Smad3 protein at the pS423 and pS425 residues. 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.
Isotype
IgG
Clonality
Polyclonal
Host
Rabbit
Gene
SMAD3
Purity
Immunogen affinity purified
Innovator's Reward
Test in a species/application not listed above to receive a full credit towards a future purchase.

Applications/Dilutions

Dilutions
  • ELISA 1:15000-1:30000
  • Flow Cytometry 1:10-1:1000
  • Immunocytochemistry/Immunofluorescence
  • Immunohistochemistry 1:500
  • Immunohistochemistry-Paraffin 1:10-1:500
  • Single Cell Western 100 ug/ml
  • Western Blot 1:2000-1:20000
Application Notes
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.
Reviewed Applications
Read 1 Review rated 3
using
NBP1-77836 in the following applications:

Publications
Read Publications using
NBP1-77836 in the following applications:

  • 2 publications
  • IHC
    1 publication
  • WB
    1 publication

Reactivity Notes

A BLAST analysis was used to suggest cross reactivity with Smad3 from Xenopus laevis, Xenopus tropicalis, zebrafish, swine, bovine and chicken based on 100% sequence homology with the immunogen. Reactivity against homologues from other sources is not known

Packaging, Storage & Formulations

Storage
Store at -20C short term. Aliquot and store at -80C long term. Avoid freeze-thaw cycles.
Buffer
0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
Preservative
0.01% Sodium Azide
Purity
Immunogen affinity purified

Notes

This affinity-purified antibody is directed against the phosphorylated form of human Smad3 protein at the [p Ser423] and [p Ser425] 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

Store vial at -20C prior to opening. Aliquot contents and freeze at -20C 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 4C as an undiluted liquid. Dilute only prior to immediate use.

Alternate Names for Smad3 [p Ser423, p Ser425] Antibody

  • DKFZp586N0721
  • DKFZp686J10186
  • hMAD-3
  • hSMAD3
  • HsT17436
  • JV15-2MGC60396
  • MAD homolog 3
  • mad protein homolog
  • MAD, mothers against decapentaplegic homolog 3 (Drosophila)
  • MAD, mothers against decapentaplegic homolog 3
  • mad3
  • MADH3mad homolog JV15-2
  • mothers against decapentaplegic homolog 3
  • Mothers against DPP homolog 3
  • pSMAD
  • SMA- and MAD-related protein 3
  • SMAD 3
  • SMAD family member 3HSPC193
  • SMAD, mothers against DPP homolog 3 (Drosophila)
  • SMAD, mothers against DPP homolog 3
  • Smad3

Background

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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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Publications for Smad3 Antibody (NBP1-77836)(6)

We have publications tested in 2 confirmed species: Human, Mouse.

We have publications tested in 3 applications: ICC/IF, IHC, WB.


Filter By Application
ICC/IF
(2)
IHC
(1)
WB
(1)
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Human
(2)
Mouse
(2)
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Review for Smad3 Antibody (NBP1-77836) (1) 31

Average Rating: 3
(Based on 1 review)
We have 1 review tested in 1 species: Feline.

Reviews using NBP1-77836:
Filter by Applications
IHC-Fr
(1)
All Applications
Filter by Species
Feline
(1)
All Species
Images Ratings Applications Species Date Details
  3
reviewed by:
Kazuya Oikawa
IHC-Fr Feline 08/15/2017
View

Summary

ApplicationImmunohistochemistry-Frozen
Sample TestedRetina
SpeciesFeline
Lot33559

Product General Protocols

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

Video Protocols

WB Video Protocol
ICC/IF Video Protocol

FAQs for Smad3 Antibody (NBP1-77836). (Showing 1 - 1 of 1 FAQs).

  1. I was wondering if you can recommend any primary polyclonal Smad3 antibody (reactivity: human) for gel supershifts?
    • We do provide a wide range of Smad3 antibodies, but unfortunately none of them have been tested for gel shift assay, so we can't guarantee they will work for an application never tested before.

Secondary Antibodies

 

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Recent Reviews

3
5
0
4
0
3
1
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1
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Kazuya Oikawa
08/15/2017
Application: IHC-Fr
Species: Feline

Bioinformatics

Gene Symbol SMAD3
Uniprot