Recombinant Anti-NCE2/UBE2F antibody [EPR12932] (ab185234)
![Western blot - Anti-NCE2/UBE2F antibody [EPR12932] (ab185234)](https://www.abcam.com/ps/products/185/ab185234/Images/ab185234-393559-anti-nce2ube2f-antibody-epr12932-western-blot-human-knockout-lysate.jpg "Western blot - Anti-NCE2/UBE2F antibody [EPR12932] (ab185234)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR12932] to NCE2/UBE2F
- Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-NCE2/UBE2F antibody [EPR12932]
See all NCE2/UBE2F primary antibodies -
Description
Rabbit monoclonal [EPR12932] to NCE2/UBE2F -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IFmore details -
Species reactivity
Reacts with: Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HeLa, Jurkat, A375 and NTERA2/D1 cell lysates. Flow Cyt (intra): A375 cells. ICC/IF: A375 cells. IHC-P: Human brain and Human renal carcinoma tissues.
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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 +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR12932 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Conjugation kits
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab185234 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) |
1/160.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
|
| WB |
1/1000. Detects a band of approximately 21 kDa (predicted molecular weight: 21 kDa).
|
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| IHC-P |
1/50 - 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
|
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| ICC/IF |
1/250.
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| Notes |
|---|
|
Flow Cyt (Intra)
1/160. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
|
WB
1/1000. Detects a band of approximately 21 kDa (predicted molecular weight: 21 kDa). |
|
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
|
ICC/IF
1/250. |
Target
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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 RBX2, but not RBX1, suggests that the RBX2-UBE2F complex neddylates specific target proteins, such as CUL5. -
Tissue specificity
Widely expressed (at protein level). -
Pathway
Protein modification; protein neddylation. -
Sequence similarities
Belongs to the ubiquitin-conjugating enzyme family. UBE2F subfamily. - Information by UniProt
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Database links
- Entrez Gene: 140739 Human
- SwissProt: Q969M7 Human
- Unigene: 471785 Human
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Alternative names
- NCE2 antibody
- NEDD8 carrier protein UBE2F antibody
- NEDD8 conjugating enzyme antibody
see all
Images
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Western blot - Anti-NCE2/UBE2F antibody [EPR12932] (ab185234)All lanes : Anti-NCE2/UBE2F antibody [EPR12932] (ab185234) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : UBE2F knockout HeLa cell lysate
Lane 3 : Jurkat 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: 21 kDa
Observed band size: 24 kDa why is the actual band size different from the predicted?Lanes 1-3: Merged signal (red and green). Green - ab185234 observed at 24 kDa. Red - loading control ab8245 observed at 36 kDa.
ab185234 Anti-NCE2/UBE2F antibody [EPR12932] was shown to specifically react with NCE2/UBE2F in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265339 (knockout cell lysate ab257777) was used. Wild-type and NCE2/UBE2F knockout samples were subjected to SDS-PAGE. ab185234 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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![Flow Cytometry (Intracellular) - Anti-NCE2/UBE2F antibody [EPR12932] (ab185234)](https://www.abcam.com/ps/products/185/ab185234/Images/ab185234-218606-anti-nce2-antibody-epr12932-flow-cytometry.jpg)
Flow Cytometry (Intracellular) - Anti-NCE2/UBE2F antibody [EPR12932] (ab185234)Intracellular flow cytometric analysis of A375 cells (2% paraformaldehyde-fixed) labeling NCE2/UBE2F with ab185234at 1/160 dilution (red) or a rabbit IgG (negative) (green), followed by Goat anti rabbit IgG (FITC) secondary at 1/150 dilution.
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![Immunocytochemistry/ Immunofluorescence - Anti-NCE2/UBE2F antibody [EPR12932] (ab185234)](https://www.abcam.com/ps/products/185/ab185234/Images/ab185234-218605-anti-nce2-antibody-epr12932-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-NCE2/UBE2F antibody [EPR12932] (ab185234)Immunofluorescent analysis of A375 cells (4% paraformaldehyde-fixed) labeling NCE2/UBE2F with ab185234 at 1/250 dilution followed by Goat anti rabbit IgG (AlexaFluor® 555) secondary at 1/200 dilution and counter-stained with DAPI (blue).
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![Western blot - Anti-NCE2/UBE2F antibody [EPR12932] (ab185234)](https://www.abcam.com/ps/products/185/ab185234/Images/ab185234-218602-anti-nce2-antibody-epr12932-western-blot.jpg)
Western blot - Anti-NCE2/UBE2F antibody [EPR12932] (ab185234)All lanes : Anti-NCE2/UBE2F antibody [EPR12932] (ab185234) at 1/1000 dilution
Lane 1 : Jurkat cell lysate
Lane 2 : A375 cell lysate
Lane 3 : NTERA2/D1 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution
Predicted band size: 21 kDa -
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NCE2/UBE2F antibody [EPR12932] (ab185234)](https://www.abcam.com/ps/products/185/ab185234/Images/ab185234-218604-anti-nce2-antibody-epr12932-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NCE2/UBE2F antibody [EPR12932] (ab185234)Immunohistochemical analysis of paraffin-embedded Human renal carcinoma labeling NCE2/UBE2F with ab185234 at 1/100 dilution followed by pre-diluted HRP-conjugated secondary antibody and counter-stained with Hematoxylin.
Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NCE2/UBE2F antibody [EPR12932] (ab185234)](https://www.abcam.com/ps/products/185/ab185234/Images/ab185234-218603-anti-nce2-antibody-epr12932-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NCE2/UBE2F antibody [EPR12932] (ab185234)Immunohistochemical analysis of paraffin-embedded Human brain labeling NCE2/UBE2F with ab185234 at 1/100 dilution followed by pre-diluted HRP-conjugated secondary antibody and counter-stained with Hematoxylin.
Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.
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![Anti-NCE2/UBE2F antibody [EPR12932] (ab185234)](https://www.abcam.com/ps/products/185/ab185234/Images/ab185234-7-benefits-of-recombinant-antibodies.png)
Anti-NCE2/UBE2F antibody [EPR12932] (ab185234)
Protocols
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (4)
ab185234 has been referenced in 4 publications.
- Guo Z et al. HUWE1-dependent DNA-PKcs neddylation modulates its autophosphorylation in DNA damage response. Cell Death Dis 11:400 (2020). PubMed: 32457294
- 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
- Kabir S et al. The CUL5 ubiquitin ligase complex mediates resistance to CDK9 and MCL1 inhibitors in lung cancer cells. Elife 8:N/A (2019). PubMed: 31294695
- 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
