Human UBE2C knockout HeLa cell line
- Advanced Validation
- What is this?
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- WB
Lab
Western blot - Human UBE2C knockout HeLa cell line (AB265032)
False colour image of Western blot : Anti-UBE2C antibody [EPR23165-31] staining at 1/1000 dilution shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution shown in red. In Western blot ab252940 was shown to bind specifically to UBE2C. A band was observed at 20 kDa in wild-type HeLa cell lysates with no signal observed at this size in UBE2C knockout cell line ab265032 (knockout cell lysate ab257775). To generate this image wild-type and UBE2C knockout HeLa cell lysates were analysed. First samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T incubated with secondary antibodies for 1 h at room temperature washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
All lanes:
Western blot - Anti-UBE2C antibody [EPR23165-31] (<a href='/en-us/products/primary-antibodies/ube2c-antibody-epr23165-31-ab252940'>ab252940</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
UBE2C knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human UBE2C knockout HeLa cell line (ab265032)
Lane 3:
WI-38 cell lysate at 20 µg
Lane 4:
K-562 (Human chronic myelogenous leukemia lymphoblast cell line ) whole cell lysate at 20 µg
Lane 5:
SW480 cell lysate at 20 µg
Lane 6:
Caco-2 cell lysate at 20 µg
Predicted band size: 20 kDa
Observed band size: 20 kDa
false
- WB
Lab
Western blot - Human UBE2C knockout HeLa cell line (AB265032)
False colour image of Western blot : Anti-UBE2C antibody staining at 1/1000 dilution shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution shown in red. In Western blot ab12290 was shown to bind specifically to UBE2C. A band was observed at 20 kDa in wild-type HeLa cell lysates with no signal observed at this size in UBE2C knockout cell line ab265032 (knockout cell lysate ab257775). To generate this image wild-type and UBE2C knockout HeLa cell lysates were analysed. First samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4 °. Blots were washed four times in TBS-T incubated with secondary antibodies for 1 h at room temperature washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
All lanes:
Western blot - Anti-UBE2C antibody (<a href='/en-us/products/primary-antibodies/ube2c-antibody-ab12290'>ab12290</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
UBE2C knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human UBE2C knockout HeLa cell line (ab265032)
Lane 3:
WI-38 cell lysate at 20 µg
Lane 4:
K-562 (Human chronic myelogenous leukemia lymphoblast cell line ) whole cell lysate at 20 µg
Lane 5:
SW480 cell lysate at 20 µg
Lane 6:
CACO2 cell lysate at 20 µg
Predicted band size: 20 kDa
Observed band size: 20 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human UBE2C knockout HeLa cell line (AB265032)
Homozygous : 1 bp deletion in exon 2.
Complementary Products
Primary Antibodies
AB252940
Anti-UBE2C antibody [EPR23165-31]
BOND RX™ Validated|RabMAb|Recombinant|KO Validated
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-UBE2C antibody [EPR23165-31] (AB252940)](https://content.abcam.com/products/images/ube2c-antibody-epr23165-31-ab252940--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img77913.jpg)
- 0
- 0
Reactivity data
Product details
We will provide viable cells that proliferate on revival.
This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant 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
Zygosity
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
UBE2C is involved in cell cycle regulation by targeting specific proteins for ubiquitination and degradation. It collaborates with the anaphase-promoting complex/cyclosome (APC/C) an important E3 ligase complex ensuring the proper progression of the cell cycle especially during the metaphase-anaphase transition. Through this action UBE2C aids in maintaining genomic stability by preventing the accumulation of proteins that can interfere with cell division.
Pathways
The role of UBE2C is integral in the cell cycle and ubiquitin-proteasome pathways. The enzyme participates in the proper execution of the metaphase-anaphase transition and degradation of cell cycle regulators like cyclin B. UBE2C interacts with proteins such as CDC20 a co-activator of APC/C to ensure accurate coordination of cell division events. Its function in the ubiquitin-proteasome pathway emphasizes its role in maintaining protein homeostasis.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
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