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RabMAb

Anti-SMAD5 抗体 [EP619Y] (ab40771)

製品の概要

  • 製品名
    Anti-SMAD5 antibody [EP619Y]
    SMAD5 一次抗体 製品一覧
  • 製品の詳細
    Rabbit monoclonal [EP619Y] to SMAD5
  • アプリケーション
    適用あり: IHC-P, WB, Flow Cyt, ICC/IFmore details
  • 種交差性
    交差種: Rat, Human, African green monkey
    交差が予測される動物種: Mouse
  • 免疫原

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human SMAD5 aa 200-300.

  • ポジティブ・コントロール
    • WB: HEK293 and Cos-1 whole cell lysate. Flow Cyt: PC-12 and HEK293 cells. ICC/IF: HeLa cells. IHC-P: Human testis tissue and human skin tissue.
  • 特記事項

    This product is a recombinant rabbit monoclonal antibody.

     

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated ‘PUR’ on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab40771 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
IHC-P 1/50.
WB 1/1000 - 1/5000. Detects a band of approximately 52 kDa (predicted molecular weight: 52 kDa).
Flow Cyt 1/30 - 1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF 1/50 - 1/100.

ターゲット情報

  • 機能
    Transcriptional modulator activated by BMP (bone morphogenetic proteins) type 1 receptor kinase. SMAD5 is a receptor-regulated SMAD (R-SMAD).
  • 組織特異性
    Ubiquitous.
  • 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 serine by BMP (bone morphogenetic proteins) type 1 receptor kinase.
    Ubiquitin-mediated proteolysis by SMAD-specific E3 ubiquitin ligase SMURF1.
  • 細胞内局在
    Cytoplasm. Nucleus. Cytoplasmic in the absence of ligand. Migrates to the nucleus when complexed with SMAD4.
  • Information by UniProt
  • 参照データベース
  • 別名
    • 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 [EP619Y] 画像

  • All lanes : Anti-SMAD5 antibody [EP619Y] (ab40771) at 1/5000 dilution (purified)

    Lane 1 : HEK293 whole cell lysate
    Lane 2 : COS-1 whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (HRP goat anti-rabbit IgG (H+L))

    Predicted band size : 52 kDa
    Observed band size : 52 kDa

    Blocking buffer: 5% NFDM/TBST
    Dilution buffer: 5% NFDM/TBST

  • Overlay histogram showing PC-12 cells fixed in 4% PFA and stained with purified ab40771 at a dilution of 1/100 (red line). The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit at a dilution of 1/500. Rabbit monoclonal IgG was used as an isotype control (black line) and cells incubated in the absence of both primary and secondary antibody were used as a negative control (blue line).

  • Immunofluorescence staining of HeLa cells with purified ab40771 at a working dilution of 1/100, counter-stained with DAPI. The secondary antibody was Alexa Fluor® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab40771 was used at a dilution of 1/500 followed by an Alexa Fluor® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.

  • Immunohistochemical staining of paraffin embedded human testis with purified ab40771 at a working dilution of 1/50. The secondary antibody used is ab97051, a goat anti-rabbit IgG (H&L) at a dilution of 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
  • Anti-SMAD5 antibody [EP619Y] (ab40771) at 1/1000 dilution (unpurified) + Cos-1 cell lysate at 10 µg

    Predicted band size : 52 kDa
    Observed band size : 52 kDa
  • Unpurified ab40771 (4µg/ml) staining SMAD5 in human skin using an automated system (DAKO Autostainer Plus). Using this protocol there is strong staining of nuclear/cytoplasmic compartments within the stratum granulosum.
    Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer EDTA pH 9.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.

  • Overlay histogram showing HEK293 cells stained with unpurified ab40771 (red line). The cells were fixed with methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab40771, 1/50 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit monoclonal IgG (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HEK293 cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

Anti-SMAD5 antibody [EP619Y] (ab40771) 使用論文

This product has been referenced in:
  • Liu P  et al. MicroRNA-23a regulates epithelial-to-mesenchymal transition in endometrial endometrioid adenocarcinoma by targeting SMAD3. Cancer Cell Int 16:67 (2016). WB . Read more (PubMed: 27601936) »
  • Dexheimer V  et al. Differential expression of TGF-ß superfamily members and role of Smad1/5/9-signalling in chondral versus endochondral chondrocyte differentiation. Sci Rep 6:36655 (2016). WB ; Human . Read more (PubMed: 27848974) »

See all 11 Publications for this product

Product Wall

Application
Western blot
Sample
Mouse Cell lysate - whole cell (liver)
Gel Running Conditions
Reduced Denaturing (4-20)
Loading amount
100 µg
Specification
liver
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

投稿 May 01 2015

Merci de nous avoir contactés.

La concentration exactedu lot de ab40771 que nous avons actuellement en stock (lot GR7392-3) est : 0.264 mg/ml.



J'espère que cette information vous sera utile. N'hésitez...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - nuclear (hESCs)
Loading amount
20 µg
Specification
hESCs
Treatment
BMP4 24h
Gel Running Conditions
Reduced Denaturing (4-12%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: 23°C
Username

Abcam user community

Verified customer

投稿 Sep 02 2008

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

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