Recombinant Anti-UBE2M/UBC12 antibody [EPR5333] (ab109507)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR5333] to UBE2M/UBC12
- Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF
- Reacts with: Human
Related conjugates and formulations
Overview
-
Product name
Anti-UBE2M/UBC12 antibody [EPR5333]
See all UBE2M/UBC12 primary antibodies -
Description
Rabbit monoclonal [EPR5333] to UBE2M/UBC12 -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IFmore details -
Species reactivity
Reacts with: Human
Predicted to work with: Mouse, Rat -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
-
Positive control
- WB: HEK-293T and Ramos cell lysates. IHC-P: Human colonic adenocarcinoma and lung carcinoma tissues. ICC/IF: Ramos cells.
-
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
-
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: 0.05% Sodium azide
Constituents: 0.1% BSA, 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant -
Concentration information loading...
-
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR5333 -
Isotype
IgG -
Research areas
Associated products
-
Alternative Versions
-
Compatible Secondaries
-
Isotype control
-
Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab109507 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 |
---|---|---|
Flow Cyt (Intra) |
1/100 - 1/1000.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
|
WB |
1/10000 - 1/50000. Detects a band of approximately 20 kDa (predicted molecular weight: 21 kDa).
|
|
IHC-P |
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
|
|
ICC/IF |
1/250.
|
Notes |
---|
Flow Cyt (Intra)
1/100 - 1/1000. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
WB
1/10000 - 1/50000. Detects a band of approximately 20 kDa (predicted molecular weight: 21 kDa). |
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
ICC/IF
1/250. |
Target
-
Function
Accepts the ubiquitin-like protein NEDD8 from the UBA3-NAE1 E1 complex and catalyzes its covalent attachment to other proteins. The specific interaction with the E3 ubiquitin ligase RBX1, but not RBX2, suggests that the RBX1-UBE2M complex neddylates specific target proteins, such as CUL1, CUL2, CUL3 and CUL4. Involved in cell proliferation. -
Pathway
Protein modification; protein neddylation. -
Sequence similarities
Belongs to the ubiquitin-conjugating enzyme family. UBC12 subfamily. -
Domain
Both the N-terminal docking peptide and the catalytic core domain must bind the UBA3-NAE1 complex simultaneously for optimal transfer of NEDD8. - Information by UniProt
-
Database links
- Entrez Gene: 9040 Human
- Entrez Gene: 22192 Mouse
- Entrez Gene: 361509 Rat
- Omim: 603173 Human
- SwissProt: P61081 Human
- SwissProt: P61082 Mouse
- Unigene: 406068 Human
- Unigene: 196580 Mouse
-
Alternative names
- hUbc12 antibody
- NEDD8 carrier protein antibody
- NEDD8 conjugating enzyme Ubc12 antibody
see all
Images
-
All lanes : Anti-UBE2M/UBC12 antibody [EPR5333] (ab109507) at 1/10000 dilution
Lane 1 : HEK-293T cell lysate
Lane 2 : Ramos cell lysate
Lysates/proteins at 10 µg per lane.
Predicted band size: 21 kDa
Observed band size: 20 kDa why is the actual band size different from the predicted? -
Immunohistochemical analysis of paraffin-embedded human colonic adenocarcinoma tissue using ab109507 at a dilution of 1/100.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
-
Immunocytochemistry/ Immunofluorescence analysis of Ramos (human Burkitt's lymphoma B lymphocyte) cells labeling UBE2M/UBC12 with purified ab109507 at 1/250 dilution (8 µg/mL). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/mL) dilution. DAPI (blue) was used as nuclear counterstain.
-
Overlay histogram showing Ramos cells stained with ab109507 (red line). The cells were fixed with 4% paraformaldehyde (10 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 (ab109507, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in Ramos cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
-
Immunohistochemical analysis of paraffin-embedded Human lung carcinoma tissue using ab109507 at a dilution of 1/100.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Protocols
Datasheets and documents
-
SDS download
-
Datasheet download
References (10)
ab109507 has been referenced in 10 publications.
- Ba C et al. Ubiquitin conjugating enzyme E2 M promotes apoptosis in osteoarthritis chondrocytes via Wnt/ß-catenin signaling. Biochem Biophys Res Commun 529:970-976 (2020). PubMed: 32819607
- Zhou L et al. Induction of NEDD8-conjugating enzyme E2 UBE2F by platinum protects lung cancer cells from apoptosis and confers to platinum-insensitivity. Cell Death Dis 11:975 (2020). PubMed: 33184273
- Guo Z et al. HUWE1-dependent DNA-PKcs neddylation modulates its autophosphorylation in DNA damage response. Cell Death Dis 11:400 (2020). PubMed: 32457294
- Zhao B et al. Knockdown of Nedd8-conjugating enzyme UBE2M suppresses the proliferation and induces the apoptosis of intrahepatic cholangiocarcinoma cells. Oncol Rep 42:2670-2679 (2019). PubMed: 31545502
- Zhou W et al. Potent 5-Cyano-6-phenyl-pyrimidin-Based Derivatives Targeting DCN1-UBE2M Interaction. J Med Chem 62:5382-5403 (2019). PubMed: 31157974
- Brockmann MM et al. Neddylation regulates excitatory synaptic transmission and plasticity. Sci Rep 9:17935 (2019). PubMed: 31784571
- Zeng Y et al. An overactive neddylation pathway serves as a therapeutic target and MLN4924 enhances the anticancer activity of cisplatin in pancreatic cancer. Oncol Lett 18:2724-2732 (2019). PubMed: 31404297
- Zhou W et al. UBE2M Is a Stress-Inducible Dual E2 for Neddylation and Ubiquitylation that Promotes Targeted Degradation of UBE2F. Mol Cell 70:1008-1024.e6 (2018). PubMed: 29932898
- Coleman KE et al. SENP8 limits aberrant neddylation of NEDD8 pathway components to promote cullin-RING ubiquitin ligase function. Elife 6:N/A (2017). PubMed: 28475037
- Xu J et al. The neddylation-cullin 2-RBX1 E3 ligase axis targets tumor suppressor RhoB for degradation in liver cancer. Mol Cell Proteomics 14:499-509 (2015). PubMed: 25540389