Anti-SMURF 2 antibody (ab94483)
- Datasheet
- References (1)
- Protocols
Overview
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Product name
Anti-SMURF 2 antibody
See all SMURF 2 primary antibodies -
Description
Rabbit polyclonal to SMURF 2 -
Host species
Rabbit -
Tested applications
Suitable for: ICC/IF, WBmore details -
Species reactivity
Reacts with: Human
Predicted to work with: Mouse, Rat, Chicken, Cow, Dog, Zebrafish
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Immunogen
Synthetic peptide corresponding to Human SMURF 2 aa 200-300 conjugated to keyhole limpet haemocyanin.
(Peptide available asab109647) -
Positive control
- This antibody gave a positive signal in the following whole cell lysates: HeLa; HepG2; Caco2; MCF7; SHSY5Y; U20S; A549.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4 -
Concentration information loading... -
Purity
Immunogen affinity purified -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
Our Abpromise guarantee covers the use of ab94483 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 |
|---|---|---|
| ICC/IF | Use a concentration of 5 µg/ml. | |
| WB | Use a concentration of 1 µg/ml. 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: 427809 Chicken
- Entrez Gene: 64750 Human
- Entrez Gene: 66313 Mouse
- Entrez Gene: 303614 Rat
- Entrez Gene: 563633 Zebrafish
- Omim: 605532 Human
- SwissProt: Q9HAU4 Human
- SwissProt: A2A5Z6 Mouse
see all -
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 (ab94483)Anti-SMURF 2 antibody (ab94483) at 1 µg/ml + U2OS (Human osteosarcoma cell line) Whole Cell Lysate at 10 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 86 kDa
Observed band size: 86 kDa
Additional bands at: 25 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 3 minutes -
Immunocytochemistry/ Immunofluorescence - Anti-SMURF 2 antibody (ab94483)ICC/IF image of ab94483 stained HepG2 cells. The cells were 4% PFA 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 (ab94483, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 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. This antibody also gave a positive result in 100% methanol fixed (5 min) HepG2 cells at 5µg/ml.
Datasheets and documents
References
This product has been referenced in:
- Li J et al. TRAF4 positively regulates the osteogenic differentiation of mesenchymal stem cells by acting as an E3 ubiquitin ligase to degrade Smurf2. Cell Death Differ 26:2652-2666 (2019). Read more (PubMed: 31076633) »
