Anti-Smad2 (phospho S467) antibody (ab53100)

Overview

  • Product nameAnti-Smad2 (phospho S467) antibodySee all Smad2 primary antibodies ...
  • Description
    Rabbit polyclonal to Smad2 (phospho S467)
  • SpecificityDetects endogenous levels of Smad2 only when phosphorylated at serine 467
  • Tested applicationsWB, ELISA more details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (Human) around the phosphorylation site of serine 467 (CSSMSP)

  • Positive control
    • HeLa cells treated with PMA

Properties

Applications

Our Abpromise guarantee covers the use of ab53100 in the following tested applications.

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

Application Notes
WB 1/300 - 1/1000. Detects a band of approximately 58 kDa (predicted molecular weight: 58 kDa).
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
  • Alternative names
    • hMAD 2 antibody
    • hMAD-2 antibody
    • hSMAD2 antibody
    • JV18 1 antibody
    • JV18 antibody
    • JV18-1 antibody
    • JV181 antibody
    • MAD antibody
    • MAD homolog 2 antibody
    • MAD Related Protein 2 antibody
    • Mad-related protein 2 antibody
    • MADH2 antibody
    • MADR2 antibody
    • MGC22139 antibody
    • MGC34440 antibody
    • Mother against DPP homolog 2 antibody
    • Mothers Against Decapentaplegic Homolog 2 antibody
    • mothers against DPP homolog 2 antibody
    • OTTHUMP00000163489 antibody
    • Sma and Mad related protein 2 antibody
    • SMAD 2 antibody
    • SMAD antibody
    • SMAD family member 2 antibody
    • SMAD, mothers against DPP homolog 2 antibody
    • SMAD2 antibody
    • SMAD2_HUMAN antibody
    see all

Anti-Smad2 (phospho S467) antibody images

  • All lanes : Anti-Smad2 (phospho S467) antibody (ab53100) at 1/300 dilution

    Lane 1 : HeLa cell extract treated with PMA (125ng/ml, 15 mins)
    Lane 2 : HeLa cell extract treated with PMA (125ng/ml, 15 mins) with phosphopeptide


    Predicted band size : 58 kDa
    Observed band size : 58 kDa
  • Anti-Smad2 (phospho S467) antibody (ab53100) at 1/750 dilution (for 16 hours at 4°C) + Mouse brain whole tissue lysate at 18 µg

    Secondary
    An HRP-conjugated Goat anti-rabbit-IgG polyclonal at 1/7000 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

  • ab53100 detecting phospho-Smad2 in Mouse brain samples by indirect ELISA. Samples were blocked with 5% BSA for 1 hour at 20°C and incubated with the primary antibody (1/750) for 4 hours at 20°C. An HRP-conjugated Goat anti-rabbit IgG polyclonal (1/5000) was used as the secondary antibody.

    See Abreview

References for Anti-Smad2 (phospho S467) antibody (ab53100)

This product has been referenced in:
  • Chiao YA  et al. Matrix metalloproteinase-9 deletion attenuates myocardial fibrosis and diastolic dysfunction in ageing mice. Cardiovasc Res 96:444-55 (2012). WB ; Mouse . Read more (PubMed: 22918978) »
  • Ivanovska N & Dimitrova P Bone resorption and remodeling in murine collagenase-induced osteoarthritis after administration of glucosamine. Arthritis Res Ther 13:R44 (2011). IHC-P ; Mouse . Read more (PubMed: 21410959) »

See all 4 Publications for this product

Product Wall

Application Western blot
Loading amount 60 µg
Gel Running Conditions Reduced Non-Denaturing (Native) (12)
Sample Human Cell lysate - nuclear (COV434 cell line)
Specification COV434 cell line
Treatment Activin A 10 ng/ml for 24 hours
Blocking step Odyssey Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 26°C
Username

Mr. FRANCESCO ELIA MARINO

Verified customer

Submitted Mar 21 2014

I can confirm that PMA is indeed Phorbol 12-myristate 13-acetate and it was used to induce the modification of Phospho-Smad2 at S467 site.

I am sorry to hear that this product has not been performing as stated on the datasheet and for the inconvenience that this has caused. This product is highly modified post translationally. Have you performed these blots under reducing conditions? T...

Read More

Thank you very much for your reply. My colleague is out of the office this week, but I'll be happy to help you with this enquiry.

You could use ab53100 in a sandwich ELISA to study the SMAD2 protein phosphorylated at residue S467. I'd suggest...

Read More

Thank you for contacting us and sorry for the delay in getting back to you.
The blocking peptide for the Anti-Smad2 (phospho S467) antibody (ab53100) is now available to purchase under the catalogue number ab138919.
I hope this information ha...

Read More

Thank you for contacting us. Because we carry over 70,000 products, it isn't feasible for us to keep small sample sizes of our products.

We are happy to reassure our customers that all of our products are covered by our Abpromise, which gu...

Read More

Thank you for contacting us.
Your credit note ID is *******.
I am sorry that the free of charge replacement (different lot) did not work.
I have asked our Finance department to issue a credit note for you against the original order # ****...

Read More

Thank you for taking time to complete our questionnaire and for contacting us. I am sorry to hear this antibody is not providing satisfactory results.
The details provided will enable us to investigate this case and will provide us with vital info...

Read More

Thank you for kindly confirming these details as this enables us to closely monitor the quality of our products
I can confirm that one vial of ab53100 with lot number is GR54038 has been added in order 1074322.
Thank you for your help and coo...

Read More

Thank you for contacting us. I am sorry to hear this antibody is not providing satisfactory results.
The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality. I apprecia...

Read More

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"