Human UBE2F (NCE2) knockout HeLa cell line
- Advanced Validation
- What is this?
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UBE2F KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 10 bp deletion in exon 2 and 187 bp insertion in exon 2.
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 line (AB265339)
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° 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 (ab265339)
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 line (AB265339)
Allele-1 : 10 bp deletion in exon 2.
- Sanger seq
Unknown
Sanger Sequencing - Human UBE2F (NCE2) knockout HeLa cell line (AB265339)
Allele-2 : 187 bp insertion in exon 2.
Reactivity data
Product details
Recommended control: Human wild-type HeLa cell line (ab255928). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.
We will provide viable cells that proliferate on revival.
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
Handling procedures
Initial handling guidelines
Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.
1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.
Subculture guidelines
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 2x104 cells/cm2 is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
Culture medium
DMEM (High Glucose) + 10% FBS
Cryopreservation medium
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
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
Target data
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'.
For licensing inquiries, please contact partnerships@abcam.com