Recombinant Anti-SMURF 2 antibody [EP629Y3] (ab53316)
![Western blot - Anti-SMURF 2 antibody [EP629Y3] (ab53316)](https://www.abcam.com/ps/products/53/ab53316/Images/ab53316-395417-anti-smurf-2-antibody-ep629y3-western-blot-human-knockout-lysate.jpg "Western blot - Anti-SMURF 2 antibody [EP629Y3] (ab53316)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP629Y3] to SMURF 2
- Suitable for: WB
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-SMURF 2 antibody [EP629Y3]
See all SMURF 2 primary antibodies -
Description
Rabbit monoclonal [EP629Y3] to SMURF 2 -
Host species
Rabbit -
Tested applications
Suitable for: WBmore details
Unsuitable for: Flow Cyt,ICC,IHC-P or IP -
Species reactivity
Reacts with: Human
Predicted to work with: Mouse, Rat
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Immunogen
Synthetic peptide within Human SMURF 2 aa 200-300. The exact sequence is proprietary.
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Positive control
- WB: HeLa, U-2 OS and SH-SY-5Y cell lysates.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 9% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA, 50% Tissue culture supernatant -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP629Y3 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab53316 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| WB | (2) |
1/500 - 1/1000. Detects a band of approximately 86 kDa (predicted molecular weight: 86 kDa).
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| Notes |
|---|
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WB
1/500 - 1/1000. Detects a band of approximately 86 kDa (predicted molecular weight: 86 kDa). |
Target
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Function
E3 ubiquitin-protein ligase which accepts ubiquitin from an E2 ubiquitin-conjugating enzyme in the form of a thioester and then directly transfers the ubiquitin to targeted substrates. Interacts with SMAD1 and SMAD7 in order to trigger their ubiquitination and proteasome-dependent degradation. In addition, interaction with SMAD7 activates autocatalytic degradation, which is prevented by interaction with SCYE1. Forms a stable complex with the TGF-beta receptor-mediated phosphorylated SMAD2 and SMAD3. In this way, SMAD2 may recruit substrates, such as SNON, for ubiquitin-mediated degradation. Enhances the inhibitory activity of SMAD7 and reduces the transcriptional activity of SMAD2. Coexpression of SMURF2 with SMAD1 results in considerable decrease in steady-state level of SMAD1 protein and a smaller decrease of SMAD2 level. -
Tissue specificity
Widely expressed. -
Pathway
Protein modification; protein ubiquitination. -
Sequence similarities
Contains 1 C2 domain.
Contains 1 HECT (E6AP-type E3 ubiquitin-protein ligase) domain.
Contains 3 WW domains. -
Domain
The second and third WW domains are responsible for interaction with the PY-motif of R-SMAD (SMAD1, SMAD2 and SMAD3).
The C2 domain is involved in autoinhibition of the catalytic activity by interacting with the HECT domain. -
Post-translational
modificationsAuto-ubiquitinated and ubiquitinated in the presence of RNF11 and UBE2D1. -
Cellular localization
Nucleus. Cytoplasm. Cell membrane. Membrane raft. Cytoplasmic in the presence of SMAD7. Co-localizes with CAV1, SMAD7 and TGF-beta receptor in membrane rafts. - Information by UniProt
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Database links
- Entrez Gene: 64750 Human
- Entrez Gene: 66313 Mouse
- Entrez Gene: 303614 Rat
- Omim: 605532 Human
- SwissProt: Q9HAU4 Human
- SwissProt: A2A5Z6 Mouse
- Unigene: 340955 Mouse
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Alternative names
- E3 ubiquitin-protein ligase SMURF2 antibody
- EC 6.3.2. antibody
- hSMURF2 antibody
see all
Images
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Western blot - Anti-SMURF 2 antibody [EP629Y3] (ab53316)All lanes : Anti-SMURF 2 antibody [EP629Y3] (ab53316) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : SMURF2 knockout HeLa cell lysate
Lane 3 : SH-SY5Y cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Predicted band size: 86 kDa
Observed band size: 86 kDaLanes 1-3: Merged signal (red and green). Green - ab53316 observed at 86 kDa. Red - loading control ab8245 observed at 36 kDa.
ab53316 Anti-SMURF 2 antibody [EP629Y3] was shown to specifically react with SMURF 2 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265231 (knockout cell lysate ab257691) was used. Wild-type and SMURF 2 knockout samples were subjected to SDS-PAGE. ab53316 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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![Western blot - Anti-SMURF 2 antibody [EP629Y3] (ab53316)](https://www.abcam.com/ps/products/53/ab53316/Images/ab53316-393638-anti-smurf-2-antibody-ep629y3-western-blot-human-knockout-lysate.jpg)
Western blot - Anti-SMURF 2 antibody [EP629Y3] (ab53316)All lanes : Anti-SMURF 2 antibody [EP629Y3] (ab53316) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : SMURF2 knockout HeLa cell lysate
Lane 3 : U-2 OS cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Predicted band size: 86 kDa
Observed band size: 86 kDaLanes 1-3: Merged signal (red and green). Green - ab53316 observed at 86 kDa. Red - loading control ab8245 observed at 36 kDa.
ab53316 Anti-SMURF 2 antibody [EP629Y3] was shown to specifically react with SMURF 2 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264806 (knockout cell lysate ab257690) was used. Wild-type and SMURF 2 knockout samples were subjected to SDS-PAGE. ab53316 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2. 5 hours at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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![Western blot - Anti-SMURF 2 antibody [EP629Y3] (ab53316)](https://www.abcam.com/ps/products/53/ab53316/Images/ab53316-66099-anti-smurf-2-antibody-ep629y3-western-blot.png)
Western blot - Anti-SMURF 2 antibody [EP629Y3] (ab53316)Anti-SMURF 2 antibody [EP629Y3] (ab53316) at 1/500 dilution + SH-SY-5Y cell lysate at 10 µg
Secondary
Goat anti-rabbit HRP labeled at 1/2000 dilution
Predicted band size: 86 kDa
Observed band size: 86 kDa -
![Anti-SMURF 2 antibody [EP629Y3] (ab53316)](https://www.abcam.com/ps/products/53/ab53316/Images/ab53316-4-benefits-of-recombinant-antibodies.png)
Anti-SMURF 2 antibody [EP629Y3] (ab53316)
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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Datasheet download
References (15)
ab53316 has been referenced in 15 publications.
- Wang Y et al. Extracellular-vesicle containing miRNA-503-5p released by macrophages contributes to atherosclerosis. Aging (Albany NY) 13:12239-12257 (2021). PubMed: 33872218
- Tang LY et al. Phosphorylation of SMURF2 by ATM exerts a negative feedback control of DNA damage response. J Biol Chem 295:18485-18493 (2020). PubMed: 33097595
- Cui Y et al. CUEDC1 inhibits epithelial-mesenchymal transition via the TßRI/Smad signaling pathway and suppresses tumor progression in non-small cell lung cancer. Aging (Albany NY) 12:20047-20068 (2020). PubMed: 33099540
- Wang R et al. Synergistic effects of miR-708-5p and miR-708-3p accelerate the progression of osteoporosis. J Int Med Res 48:300060520978015 (2020). PubMed: 33322976
- Mund T & Pelham HR Substrate clustering potently regulates the activity of WW-HECT domain-containing ubiquitin ligases. J Biol Chem 293:5200-5209 (2018). PubMed: 29463679
