Rabbit Recombinant Monoclonal NCE2/UBE2F antibody. Carrier free. Suitable for WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Human samples.
pH: 7.2 - 7.4
Constituents: PBS
WB | ICC/IF | Flow Cyt (Intra) | IHC-P | |
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Human | Tested | Tested | Tested | Tested |
Species | Dilution info | Notes |
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Species Human | Dilution info - | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
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Species Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Human | Dilution info - | Notes ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info - | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Accepts the ubiquitin-like protein NEDD8 from the UBA3-NAE1 E1 complex and catalyzes its covalent attachment to other proteins (PubMed:19250909, PubMed:23201271). Together with the E3 ubiquitin ligase RNF7/RBX2, specifically neddylates cullin-5 (CUL5) (PubMed:19250909, PubMed:23201271, PubMed:23300442). Does not neddylate CUL1, CUL2, CUL3, CUL4A or CUL4B (PubMed:19250909, PubMed:23201271). Mediates neddylation of the CUL9-RBX1 complex (PubMed:38605244). (Microbial infection) Following infection by HIV-1 virus, participates to HIV-1 Vif protein-mediated ubiquitination and degradation of APOBEC3G by mediating neddylation of cullin-5 (CUL5).
NCE2, UBE2F, NEDD8-conjugating enzyme UBE2F, NEDD8 carrier protein UBE2F, NEDD8 protein ligase UBE2F, NEDD8-conjugating enzyme 2, RING-type E3 NEDD8 transferase UBE2F, Ubiquitin-conjugating enzyme E2 F
Rabbit Recombinant Monoclonal NCE2/UBE2F antibody. Carrier free. Suitable for WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Human samples.
pH: 7.2 - 7.4
Constituents: PBS
ab240291 is the carrier-free version of Anti-NCE2/UBE2F antibody [EPR12932] ab185234.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
This product is a recombinant monoclonal antibody, which offers several advantages including:
For more information, read more on recombinant antibodies.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
NCE2/UBE2F also known simply as UBE2F acts as a ubiquitin-conjugating enzyme that is critical in the process of neddylation. The mass of UBE2F is approximately 21 kDa. It plays a mechanical role in transferring the activated NEDD8 protein to target substrates which include members of the Cullin-RING ligase complexes. Expression of UBE2F is observed in multiple human tissues with notable presence in the lungs and liver. Its activity ensures the proper modification of proteins that depend on neddylation for their function.
UBE2F functions as an important component in cellular processes by participating in the neddylation pathway. It forms a complex with NEDD8-activating E1 enzymes facilitating the transfer of NEDD8 to specific E3 ligases primarily targeting cullins. Through neddylation UBE2F influences the activation of these ligases impacting protein degradation pathways cell cycle regulation and signal transduction. This functional role positions UBE2F as an important player in maintaining cellular homeostasis.
UBE2F integrates into the neddylation and ubiquitination pathways. Neddylation specifically modulates ubiquitin-proteasome system where NEDD8 acts as a regulator of Cullin-RING E3 ligase activity. Through these interactions UBE2F contributes to the regulation of protein turnover alongside ubiquitin itself. Connections in these pathways often involve proteins like CUL5 which works synchronously with UBE2F to ensure proper cell cycle progression and stress response in cells.
UBE2F has associations with cancer development such as lung cancer and neurodegenerative conditions like Parkinson's disease. Its role in cancer ties to abnormal neddylation activity where overactive maintenance of cullin function disrupts cell cycle control. In Parkinson’s UBE2F-related dysregulation affects protein homeostasis and neuronal health. Proteins linked to disorders like UbcH12 for cancer highlight UBE2F's involvement in complex cellular mechanisms that contribute to disease pathophysiology.
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
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This data was developed using the same antibody clone in a different buffer formulation (Anti-NCE2/UBE2F antibody [EPR12932] ab185234).
Lanes 1-3: Merged signal (red and green). Green - Anti-NCE2/UBE2F antibody [EPR12932] ab185234 observed at 24 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 36 kDa.
Anti-NCE2/UBE2F antibody [EPR12932] 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 Human UBE2F (NCE2) knockout HeLa cell line ab265339 (knockout cell lysate Human UBE2F (NCE2) knockout HeLa cell lysate ab257777) was used. Wild-type and NCE2/UBE2F knockout samples were subjected to SDS-PAGE. Anti-NCE2/UBE2F antibody [EPR12932] ab185234 and Anti-GAPDH antibody [6C5] - Loading Control (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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (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.
All lanes: Western blot - Anti-NCE2/UBE2F antibody [EPR12932] (Anti-NCE2/UBE2F antibody [EPR12932] ab185234) at 1/1000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: UBE2F knockout HeLa cell lysate at 20 µg
Lane 2: Western blot - Human UBE2F (NCE2) knockout HeLa cell line (Human UBE2F (NCE2) knockout HeLa cell line ab265339)
Lane 3: Jurkat cell lysate at 20 µg
All lanes: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/10000 dilution
Predicted band size: 21 kDa
Observed band size: 24 kDa
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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-NCE2/UBE2F antibody [EPR12932] ab185234).
Immunofluorescent analysis of A375 cells (4% paraformaldehyde-fixed) labeling NCE2/UBE2F with Anti-NCE2/UBE2F antibody [EPR12932] ab185234 at 1/250 dilution followed by Goat anti rabbit IgG (AlexaFluor® 555) secondary at 1/200 dilution and counter-stained with DAPI (blue).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-NCE2/UBE2F antibody [EPR12932] ab185234).
Immunohistochemical analysis of paraffin-embedded Human renal carcinoma labeling NCE2/UBE2F with Anti-NCE2/UBE2F antibody [EPR12932] ab185234 at 1/100 dilution followed by pre-diluted HRP-conjugated secondary antibody and counter-stained with Hematoxylin.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-NCE2/UBE2F antibody [EPR12932] ab185234).
Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.
Immunohistochemical analysis of paraffin-embedded Human brain labeling NCE2/UBE2F with Anti-NCE2/UBE2F antibody [EPR12932] ab185234 at 1/100 dilution followed by pre-diluted HRP-conjugated secondary antibody and counter-stained with Hematoxylin.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-NCE2/UBE2F antibody [EPR12932] ab185234).
Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.
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