概述

  • 产品名称Anti-SMAD5抗体
    参阅全部 SMAD5 一抗
  • 描述
    兔多克隆抗体to SMAD5
  • 经测试应用适用于: WB, IP, ICC/IFmore details
  • 种属反应性
    与反应: Mouse, Rat, Human, Zebrafish
  • 免疫原

    Synthetic peptides, corresponding to amino acids 20-34 (GWKQGDEEEKWAEKA) and 316-330 (VNRNSTIENTRRHIG) of Human SMAD 5. These sequences are conserved in human Smad 1, 8 and 9 and expected to cross react due to sequence homology.

  • 阳性对照
    • Tested with Saos-2 cell lysate. This antibody gave a positive result in IHC in the following FFPE tissue: Human normal lung.

性能

应用

Our Abpromise guarantee covers the use of ab13724 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 1/500 - 1/2000. Detects a band of approximately 52 kDa (predicted molecular weight: 52 kDa).
IP Use at an assay dependent concentration.
ICC/IF 1/1000.

靶标

  • 功能Transcriptional modulator activated by BMP (bone morphogenetic proteins) type 1 receptor kinase. SMAD5 is a receptor-regulated SMAD (R-SMAD).
  • 组织特异性Ubiquitous.
  • 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 serine by BMP (bone morphogenetic proteins) type 1 receptor kinase.
    Ubiquitin-mediated proteolysis by SMAD-specific E3 ubiquitin ligase SMURF1.
  • 细胞定位Cytoplasm. Nucleus. Cytoplasmic in the absence of ligand. Migrates to the nucleus when complexed with SMAD4.
  • Information by UniProt
  • 数据库链接
  • 别名
    • DKFZp781C1895 antibody
    • DKFZp781O1323 antibody
    • Dwfc antibody
    • hSmad5 antibody
    • JV5 1 antibody
    • JV5-1 antibody
    • MAD homolog 5 antibody
    • MAD, mothers against decapentaplegic homolog 5 antibody
    • MADH 5 antibody
    • MADH5 antibody
    • Mothers against decapentaplegic homolog 5 antibody
    • mothers against decapentaplegic, drosophila, homolog of, 5 antibody
    • Mothers against DPP homolog 5 antibody
    • MusMLP antibody
    • SMA and MAD related protein 5 antibody
    • SMAD 5 antibody
    • SMAD family member 5 antibody
    • SMAD, mothers against DPP homolog 5 antibody
    • Smad5 antibody
    • SMAD5_HUMAN antibody
    see all

Anti-SMAD5 antibody 图像

  • ICC/IF image of ab13724 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab13724, 1/1000 dilution) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • All lanes : Anti-SMAD5 antibody (ab13724) at 1/500 dilution

    Lane 1 : Whole embryo lysates prepared from Zebrafish as control morpholinos.
    Lane 2 : Whole embryo lysates prepared from Zebrafish embryos injected with ints5 morpholinos.


    Predicted band size : 52 kDa

    Image from Tao Shijie et. al., Development 136, 2757-2765 (2009), Fig S4.

  • IHC image of SMAD5 staining in Human normal lung formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab13724, 1/200 dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

Anti-SMAD5 antibody (ab13724)参考文献

This product has been referenced in:
  • Lu H  et al. Hepcidin promotes osteogenic differentiation through the bone morphogenetic protein 2/small mothers against decapentaplegic and mitogen-activated protein kinase/P38 signaling pathways in mesenchymal stem cells. Mol Med Rep 11:143-50 (2015). WB ; Rat . Read more (PubMed: 25351366) »
  • Tao S  et al. The Integrator subunits function in hematopoiesis by modulating Smad/BMP signaling. Development 136:2757-65 (2009). WB ; Zebrafish . Read more (PubMed: 19605500) »

See all 3 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (human osteosarcoma saos2)
Specification human osteosarcoma saos2
Fixative Paraformaldehyde
Permeabilization Yes - PBS-TRITON 0.25%
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Username

Abcam user community

Verified customer

提交于 Apr 19 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Zebrafish Cell lysate - whole cell (whole embryo)
Loading amount 10 µg
Specification whole embryo
Gel Running Conditions Reduced Denaturing (10% PAGE gel)
Blocking step Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Username

Abcam user community

Verified customer

提交于 Jan 04 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunoprecipitation
Sample Mouse Cell lysate - whole cell (lung epithelium)
Total protein in input 600 µg
Specification lung epithelium
Immuno-precipitation step Protein A/G
Username

Abcam user community

Verified customer

提交于 May 18 2010

Thank you for your enquiry regarding ab13724. The immunogen sequences used to generate the anti-Smad5 antibody are amino acids 20-34 (GWKQGDEEEKWAEKA) and 316-330 (VNRNSTIENTRRHIG). When I used BLAST to search for the first sequence I found both Smad...

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