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Overview

  • Product nameAnti-Smad2 antibodySee all Smad2 primary antibodies ...
  • Description
    Rabbit polyclonal to Smad2
  • Tested applicationsIHC-Fr, WB, IHC-P, ICC/IF, ELISA more details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic non-phosphopeptide derived from human Smad2 around the phosphorylation site of serine 467 (C-S-S-M-S).

  • Positive controlHepG2 cells. Prostate carcinoma.

Properties

Applications

Our Abpromise guarantee covers the use of ab63576 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Notes
IHC-Fr IHC-Fr: Use at an assay dependent concentration.
WB WB: 1/300 - 1/1000. Detects a band of approximately 58 kDa (predicted molecular weight: 58 kDa).
IHC-P IHC-P: 1/50 - 1/100.
ICC/IF ICC/IF: 1/500 - 1/1000.
ELISA ELISA: 1/5000.

Target

  • FunctionReceptor-regulated SMAD (R-SMAD) that is an intracellular signal transducer and transcriptional modulator activated by TGF-beta (transforming growth factor) and activin type 1 receptor kinases. Binds the TRE element in the promoter region of many genes that are regulated by TGF-beta and, on formation of the SMAD2/SMAD4 complex, activates transcription. May act as a tumor suppressor in colorectal carcinoma.
  • Tissue specificityExpressed at high levels in skeletal muscle, heart and placenta.
  • Sequence similaritiesBelongs to the dwarfin/SMAD family.
    Contains 1 MH1 (MAD homology 1) domain.
    Contains 1 MH2 (MAD homology 2) domain.
  • Post-translational
    modifications    Phosphorylated on one or several of Thr-220, Ser-245, Ser-250, and Ser-255. In response to TGF-beta, phosphorylated on Ser-465/467 by TGF-beta and activin type 1 receptor kinases. Able to interact with SMURF2 when phosphorylated on Ser-465/467, recruiting other proteins, such as SNON, for degradation. In response to decorin, the naturally occurring inhibitor of TGF-beta signaling, phosphorylated on Ser-240 by CaMK2. Phosphorylated by MAPK3 upon EGF stimulation; which increases transcriptional activity and stability, and is blocked by calmodulin.
    In response to TGF-beta, ubiquitinated by NEDD4L; which promotes its degradation.
    Acetylated on Lys-19 by coactivators in response to TGF-beta signaling, which increases transcriptional activity. Isoform short: Acetylation increases DNA binding activity in vitro and enhances its association with target promoters in vivo. Acetylation in the nucleus by EP300 is enhanced by TGF-beta.
  • Cellular localizationCytoplasm. Nucleus. Cytoplasmic and nuclear in the absence of TGF-beta. On TGF-beta stimulation, migrates to the nucleus when complexed with SMAD4. On dephosphorylation by phosphatase PPM1A, released from the SMAD2/SMAD4 complex, and exported out of the nucleus by interaction with RANBP1.

    • Target information above from: UniProt accession Q15796 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links
    see all
  • Alternative names
      hMAD 2 antibodyhMAD-2 antibodyhSMAD2 antibody
      JV18 1 antibodyJV18 antibodyJV18-1 antibodyJV181 antibodyMAD antibodyMAD homolog 2 antibodyMAD Related Protein 2 antibodyMad-related protein 2 antibodyMADH2 antibodyMADR2 antibodyMGC22139 antibodyMGC34440 antibodyMothers Against Decapentaplegic Homolog 2 antibodymothers against DPP homolog 2 antibodyOTTHUMP00000163489 antibodySma and Mad related protein 2 antibodySMAD 2 antibodySMAD antibodySMAD family member 2 antibodySMAD, mothers against DPP homolog 2 antibodySMAD2 antibodySMAD2_HUMAN antibody
    see all

Anti-Smad2 antibody images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Smad2 antibody (ab63576)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Smad2 antibody (ab63576)
    ab63576, at a 1/50 dilution, staining human Smad2 in prostate carcinoma, using Immunohistochemistry, Formalin/PFA fixed Paraffin embedded tissue, minus immunising peptide (left image) and plus immunising peptide (right image).
  • Immunocytochemistry/ Immunofluorescence - Smad2 antibody (ab63576)
    Immunocytochemistry/ Immunofluorescence - Smad2 antibody (ab63576)
    Immunofluorescent staining of HepG2 cells, using ab63576 at a 1/500 dilution, minus immunising peptide (left image) and plus immunising peptide (right image).
  • Immunocytochemistry/ Immunofluorescence - Smad2 antibody (ab63576)
    Immunocytochemistry/ Immunofluorescence - Smad2 antibody (ab63576)This image is courtesy of an anonymous Abreview
    ab63576 staining Smad2 in Mouse enterocytes by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.5% Triton and blocked with 5% BSA for 1 hour at 20°C. Samples were incubated with primary antibody (1/250) for 2 hours at 20°C. An Alexa Fluor®488-conjugated Goat anti-rabbit IgG polyclonal (1/1000) was used as the secondary antibody.

    See Abreview

  • Western blot - Smad2 antibody (ab63576)
    Western blot - Smad2 antibody (ab63576)
    Anti-Smad2 antibody (ab63576) at 1/1000 dilution (for 17 hours at 4°C) + Mouse brain whole cell lysate at 18 µg

    Secondary
    An HRP-conjugated Goat anti-rabbit IgG polyclonal at 1/10000 dilution
    developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 58 kDa
    Observed band size : 58 kDa


    Exposure time : 2 minutes

    This image is courtesy of an anonymous Abreview

    See Abreview

  • Western blot - Smad2 antibody (ab63576)
    Western blot - Smad2 antibody (ab63576)
    Anti-Smad2 antibody (ab63576) at 1/1000 dilution (for 16 hours at 4°C) + Rat intestine whole cell lysate at 25 µg

    Secondary
    An HRP-conjugated Goat anti-rabbit IgG at 1/10000 dilution
    developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 58 kDa
    Observed band size : 58 kDa


    Exposure time : 140 seconds

    This image is courtesy of an anonymous Abreview

    See Abreview

References for Anti-Smad2 antibody (ab63576)

This product has been referenced in:
  • Spiller CM  et al. Endogenous Nodal signaling regulates germ cell potency during mammalian testis development. Development 139:4123-32 (2012). IHC-P ; Mouse . Read more (PubMed: 23034635) »

See 1 Publication for this product

Publishing research using ab63576 ? Please let us know so that we can cite the reference in this datasheet

Product Wall

Submit an Abreview for ab63576 Submit a Question for ab63576

Displaying 1 - 10 of 14 results for Abreviews and Q&A

10
Votes: Highest First

als Ersatz hätte ich gerne einen neuen Antikörper. Falls sie vom ab63576 noch einen anderen Lot haben, würde ich diesen gerne nehmen. Ansonsten einen weiteren anti-Smad2: ab119907
Bestellt wurde ursprünglich am 24.5.12
Weitere Angaben finden Sie i...

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Es tut mir leid, dass Sie Probleme mit diesem Antikörper hatten. Wie besprochen habe ich eine kostenlose Ersatzlieferung für Sie in Auftrag gegeben. Sie hat die Referenznummer 1111111 und sollte am Mittwoch bei Ihnen ankommen.

Ich wünsche Ihnen v...

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hier meine Angaben.




1) Abcam product code ab63576



2) Abcam order reference number or product batch number : GR24273-3





3) Description of the problem :

bands f...

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Es tut mir leid zu hören, dass unsere Vorschläge in diesem Fall nicht zu einer Verbesserung Ihrer Ergebnisse geführt haben. Ich weiß, dass Sie viel Zeit für diese Experimente verwendet haben und möchte Ihnen daher eine Gutschrift, einen Antikörper oder ei...

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I think first that we would simply like to get a replacement vial and test it to ensure that it wasn’t just a stability issue or something. Is that possible to do first?

Thank you for your reply.

I have sent you a new vial of ab63576 as requested. Your new order number is *********** and you should have the antibody by 10.30am tomorrow morning.

Please let me know if you have any further issues with thi...

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Thanks for following up. Here are the original purchase order and quote #:

Our PO **********

Abcam Quote #****

Thank you for your reply.

I forgot to ask in the previous email which Smad2 antibody you would like as a replacement. Below are a few that might be suitable:

http://www.abcam.com/Smad2-antibody-EP567Y-ab33875.html
http://www.abcam...

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Hi there. We have come across an issue with an antibody. We essentially got no binding/staining/results using ab63576. Here are the details:

Thank you for contacting Abcam.

The antibody is covered under our Abpromise for six months and is guaranteed to work in western blot on human samples. Based on the information you have provided, I do not think there will be anything I can sugges...

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We used the ab for WB. We had no good signal and a lot of nonspecific bands. We use Jurkat and Hela cells and compared your ab with another company's Ab (see pictures, which also show ab dilutions). The first was probed with Smad2 that we bought from ...

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Thank you for your reply and the additional protocol information. The WB image you sent - was that for the total Smad2 (ab63576) or the phospho-Smad2 (ab53100)? If it was the total Smad 2, what did the blot for phospho-Smad2 look like? Was it the sam...

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Immunohistochemistry (Frozen sections) abreview for Anti-Smad2 antibody

 Good
Abreviews
Application Immunohistochemistry (Frozen sections)
Sample Human Tissue sections (Human Intestine tissue sections)
Specification Human Intestine tissue sections
Fixative Acetone
Permeabilization No
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
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Verified customer

Submitted Sep 17 2010

Western blot abreview for Anti-Smad2 antibody

 Good
Abreviews
Application Western blot
Sample Human Tissue lysate - whole (Human Brain)
Loading amount 18 µg
Specification Human Brain
Gel Running Conditions Reduced Denaturing (10%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
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Submitted Sep 16 2010

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Smad2 antibody

 Good
Abreviews
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (Human Intestine)
Specification Human Intestine
Fixative Acetone
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citric acid pH 5.5
Permeabilization No
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 18°C
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Verified customer

Submitted Sep 03 2010

Western blot abreview for Anti-Smad2 antibody

 Good
Abreviews
Application Western blot
Sample Rat Tissue lysate - whole (Rat intestine)
Loading amount 25 µg
Specification Rat intestine
Gel Running Conditions Reduced Denaturing (10%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
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Verified customer

Submitted Aug 27 2010



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