製品の概要

  • 製品名
    Anti-Smad2 (phospho S467) antibody
    Smad2 一次抗体 製品一覧
  • 製品の詳細
    Rabbit polyclonal to Smad2 (phospho S467)
  • 特異性
    Detects endogenous levels of Smad2 only when phosphorylated at serine 467
  • アプリケーション
    適用あり: WB, ELISA, IHC-Pmore details
  • 種交差性
    交差種: Mouse, Rat, Human
  • 免疫原

    Synthetic peptide corresponding to Human Smad2.
    (Peptide available as ab138919)

  • ポジティブ・コントロール
    • HeLa cells treated with PMA

製品の特性

アプリケーション

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.

アプリケーション Abreviews 特記事項
WB 1/300 - 1/1000. Detects a band of approximately 58 kDa (predicted molecular weight: 58 kDa).
ELISA 1/5000.
IHC-P 1/50 - 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

ターゲット情報

  • 機能
    Receptor-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.
  • 組織特異性
    Expressed at high levels in skeletal muscle, heart and placenta.
  • 配列類似性
    Belongs to the dwarfin/SMAD family.
    Contains 1 MH1 (MAD homology 1) domain.
    Contains 1 MH2 (MAD homology 2) domain.
  • 翻訳後修飾
    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.
  • 細胞内局在
    Cytoplasm. 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.
  • Information by UniProt
  • 参照データベース
  • 別名
    • Drosophila, homolog of, MADR2 antibody
    • hMAD-2 antibody
    • HsMAD2 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 decapentaplegic, Drosophila, homolog of, 2 antibody
    • Mothers against DPP homolog 2 antibody
    • OTTHUMP00000163489 antibody
    • Sma and Mad related protein 2 antibody
    • Sma- and Mad-related protein 2 MAD antibody
    • SMAD 2 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 画像

  • 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
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue labeling Smad2 (phospho S467) with ab53100. The image on the right is blocked with a phosphopeptide.

     

  • 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

    Blocking Step: 5% Milk for 1 hour at 18°C

    See Abreview

  • Anti-Smad2 (phospho S467) antibody (ab53100) at 1/1500 dilution + Rat lung whole tissue lysate at 15 µg with Milk at 5 %

    Secondary
    Goat antirabbit conjugated to HRP at 1/7000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 58 kDa
    Additional bands at : 59 kDa. We are unsure as to the identity of these extra bands.

    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

Anti-Smad2 (phospho S467) antibody (ab53100) 使用論文

This product has been referenced in:
  • Mao SY  et al. The role of ZFP580, a novel zinc finger protein, in TGF-mediated cytoprotection against chemical hypoxia-induced apoptosis in H9c2 cardiac myocytes. Mol Med Rep 15:2154-2162 (2017). Read more (PubMed: 28259939) »
  • Fang L  et al. Jumonji AT-rich interactive domain 1B overexpression is associated with the development and progression of glioma. Int J Mol Med 38:172-82 (2016). Read more (PubMed: 27246838) »

See all 10 Publications for this product

Product Wall

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Lung, colon cancer, normal colon, Glioblastoma)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate, Tris EDTA
Permeabilization
No
Specification
Lung, colon cancer, normal colon, Glioblastoma
Blocking step
Milk as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative
Formaldehyde
Username

Sravya Palavalasa

Verified customer

投稿 May 16 2016

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
Odyssey Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 26°C
Sample
Human Cell (Ovarian Granulosa Cell Line)
Specification
Ovarian Granulosa Cell Line
Permeabilization
Yes - Ethanol and Triton
Fixative
Paraformaldehyde
Username

Dr. Francesco Elia Marino

Verified customer

投稿 Jun 10 2014

Application
Western blot
Loading amount
40 µg
Gel Running Conditions
Reduced Non-Denaturing (Native) (12)
Sample
Mouse Cell lysate - whole cell (Gastrocnemius)
Specification
Gastrocnemius
Blocking step
Commercial Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 26°C
Username

Abcam user community

Verified customer

投稿 May 23 2014

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

Dr. Francesco Elia Marino

Verified customer

投稿 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...

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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...

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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...

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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...

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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 # ****...

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

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