Anti-Smad2 (phospho S467)抗体(ab53100)

概述

  • 产品名称
    Anti-Smad2 (phospho S467)抗体
    参阅全部 Smad2 一抗
  • 描述
    兔多克隆抗体to Smad2 (phospho S467)
  • 宿主
    Rabbit
  • 特异性
    Detects endogenous levels of Smad2 only when phosphorylated at serine 467
  • 经测试应用
    适用于: WB, ELISA, IHC-Pmore details
  • 种属反应性
    与反应: Mouse, Rat, Human
  • 免疫原

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

  • 阳性对照
    • HeLa cells treated with PMA

性能

应用

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.

应用 Ab评论 说明
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.

靶标

  • 功能
    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.
  • 组织特异性
    Expressed at high levels in skeletal muscle, heart and placenta.
  • 序列相似性
    Belongs to the dwarfin/SMAD family.
    Contains 1 MH1 (MAD homology 1) domain.
    Contains 1 MH2 (MAD homology 2) domain.
  • 翻译后修饰
    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.
  • 细胞定位
    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
  • 数据库链接
  • 别名
    • 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

图片

  • 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


    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

    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

文献

This product has been referenced in:
  • Yang HJ  et al. GP73 promotes invasion and metastasis of bladder cancer by regulating the epithelial-mesenchymal transition through the TGF-ß1/Smad2 signalling pathway. J Cell Mol Med 22:1650-1665 (2018). WB ; Human . Read more (PubMed: 29349903) »
  • Zhang X  et al. ING5 is a Potential Target for Osteosarcoma Therapy. Technol Cancer Res Treat 17:1533033818762680 (2018). Read more (PubMed: 29528777) »

See all 23 Publications for this product

客户评价及客户问答

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Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (embryo heart)
Permeabilization
Yes - 0.3% Triton
Specification
embryo heart
Blocking step
Normal Donkey Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

提交于 Mar 07 2018

Application
Western blot
Sample
Rat Tissue lysate - whole (Muscle)
Gel Running Conditions
Reduced Denaturing (9)
Loading amount
50 µg
Specification
Muscle
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Mr. Baptiste Jude

Verified customer

提交于 Feb 26 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Prostate)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate, pH 6
Permeabilization
No
Specification
Prostate
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 20°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

提交于 Dec 29 2017

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

提交于 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

提交于 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

提交于 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

提交于 Mar 21 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Rat Tissue lysate - whole (Rat lungs)
Loading amount
15 µg
Specification
Rat lungs
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
Username

Abcam user community

Verified customer

提交于 Sep 16 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Tissue lysate - whole (Human kidney)
Loading amount
20 µg
Specification
Human kidney
Gel Running Conditions
Reduced Denaturing (10%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Username

Abcam user community

Verified customer

提交于 Sep 16 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
ELISA
Sample
Mouse Tissue sections (Mouse brain)
Specification
Mouse brain
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Type
Indirect
Username

Abcam user community

Verified customer

提交于 Aug 20 2010

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