UBE2C KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon 2.
HeLa
Human
Cervix
Liquid
Sanger Sequencing, Western blot
Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon 2
Cyclin selective ubiquitin carrier protein, Mitotic specific ubiquitin conjugating enzyme, UBCH 10, UBE2C_HUMAN, Ubiquitin carrier protein C, Ubiquitin carrier protein E2 C, Ubiquitin-conjugating enzyme E2 C, Ubiquitin-protein ligase C, dJ447F3.2
UBE2C KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon 2.
HeLa
Human
Cervix
Liquid
Sanger Sequencing, Western blot
Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon 2
Adenocarcinoma
UBE2C
Knockout
CRISPR technology
Sanger Sequencing, Western blot
Homozygous
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
EU: 2 US: 2
Adherent
Female
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.
DMEM (High Glucose) + 10% FBS
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Dry Ice
-196°C
-196°C
Recommended control: Human wild-type HeLa cell line (Human wild-type HeLa cell line ab255448). 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 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.
This supplementary information is collated from multiple sources and compiled automatically.
UBE2C also known as ubiquitin-conjugating enzyme E2C or UBCH10 is an enzyme that plays a role in the ubiquitination process. Its molecular mass is approximately 20 kDa. UBE2C is an E2 ubiquitin-conjugating enzyme actively involved in marking proteins for degradation through the ubiquitin-proteasome system. It facilitates the transfer of ubiquitin to target proteins cooperating with E3 ligases. UBE2C shows expression in various tissues with high levels observed in proliferating cells such as those found in tumors.
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.
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.
UBE2C's dysfunction or overexpression associates with cancer development and progression. Overexpression of UBE2C has been observed in various cancers including breast cancer and prostate cancer and may contribute to tumorigenesis by disrupting cell cycle control. UBE2C can interact with oncoproteins such as MYC further linking its dysregulation to cancer pathways. Understanding the relationships between UBE2C and cancer may offer insights into therapeutic targets for tumor treatment.
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False colour image of Western blot: Anti-UBE2C antibody staining at 1/1000 dilution shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution shown in red. In Western blot Anti-UBE2C antibody 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 Human UBE2C knockout HeLa 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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.
All lanes: Western blot - Anti-UBE2C antibody (Anti-UBE2C antibody ab12290) 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 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
Performed under reducing conditions.
Predicted band size: 20 kDa
Observed band size: 20 kDa
False colour image of Western blot: Anti-UBE2C antibody [EPR23165-31] staining at 1/1000 dilution shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution shown in red. In Western blot Anti-UBE2C antibody [EPR23165-31] 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 Human UBE2C knockout HeLa 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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.
All lanes: Western blot - Anti-UBE2C antibody [EPR23165-31] (Anti-UBE2C antibody [EPR23165-31] ab252940) 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 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
Performed under reducing conditions.
Predicted band size: 20 kDa
Observed band size: 20 kDa
Homozygous: 1 bp deletion in exon 2.
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