Human UBE2F (NCE2) knockout HeLa cell lysate
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UBE2F KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 10 bp deletion in exon2 and 187 bp insertion in exon2.
View Alternative Names
NCE2, NEDD8 carrier protein UBE2F, NEDD8 conjugating enzyme, NEDD8 protein ligase UBE2F, NEDD8-conjugating enzyme 2, NEDD8-conjugating enzyme UBE2F, UBE2F_HUMAN, Ubiquitin conjugating enzyme E2F (putative), Ubiquitin-conjugating enzyme E2 F
- WB
Lab
Western blot - Human UBE2F (NCE2) knockout HeLa cell lysate (AB257777)
Lane 1 : Wild-type HeLa cell lysate (20 ug)
Lane 2 : UBE2F knockout HeLa cell lysate (20 ug)
Lane 3 : Jurkat cell lysate (20 ug)
ab185234 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 4oC 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.
All lanes:
Western blot - Anti-NCE2/UBE2F antibody [EPR12932] (<a href='/en-us/products/primary-antibodies/nce2-ube2f-antibody-epr12932-ab185234'>ab185234</a>) 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 (<a href='/en-us/products/cell-lines/human-ube2f-nce2-knockout-hela-cell-line-ab265339'>ab265339</a>)
Lane 3:
Jurkat cell lysate at 20 µg
Secondary
All lanes:
Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution
Predicted band size: 21 kDa
Observed band size: 24 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human UBE2F (NCE2) knockout HeLa cell lysate (AB257777)
Allele-1 : 10 bp deletion in exon2
- Sanger seq
Unknown
Sanger Sequencing - Human UBE2F (NCE2) knockout HeLa cell lysate (AB257777)
Allele-2 : 187 bp insertion in exon2
Reactivity data
Product details
Knockout cell lysate achieved by CRISPR/Cas9.
REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.
Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.
User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.
This product is subject to limited use licenses from The Broad Institute, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
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.
Pathways
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.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Complementary Products
![Western blot - Anti-NCE2/UBE2F antibody [EPR12932] (AB185234)](https://content.abcam.com/products/images/nce2-ube2f-antibody-epr12932-ab185234--western-blot-img106382.jpg)
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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