Recombinant 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/IF,IHC-P or IP -
Species reactivity
Reacts with: Human
Predicted to work with: Mouse, Rat -
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...
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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 |
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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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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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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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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
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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SDS download
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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