UBE2A KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 19 bp deletion in exon 1 and 8 bp deletion in exon 1.
Images
Key facts
- Cell type
- HEK-293T
- Species or organism
- Human
- Tissue
- Kidney
- Form
- Liquid
- Knockout validation
- Sanger Sequencing
- Mutation description
- Knockout achieved by using CRISPR/Cas9, 19 bp deletion in exon 1 and 8 bp deletion in exon 1
Alternative names
BHR6A, HR6A, MRXS30, MRXSN, RAD6 homolog A, RAD6A, UBC2, UBCD6, UBE2A_HUMAN, Ube2a ubiquitin-conjugating enzyme E2A, Ubiquitin carrier protein, Ubiquitin carrier protein A, Ubiquitin conjugating enzyme E2 21.5 kDa, Ubiquitin conjugating enzyme E2A (RAD6 homolog), Ubiquitin-conjugating enzyme E2 A, Ubiquitin-conjugating enzyme E2-17 kDa, Ubiquitin-protein ligase A, hHR6A, mHR6A
Recommended products
UBE2A KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 19 bp deletion in exon 1 and 8 bp deletion in exon 1.
Key facts
- Cell type
- HEK-293T
- Species or organism
- Human
- Tissue
- Kidney
- Form
- Liquid
- Knockout validation
- Sanger Sequencing
- Mutation description
- Knockout achieved by using CRISPR/Cas9, 19 bp deletion in exon 1 and 8 bp deletion in exon 1
- Concentration
Properties
- Gene name
- UBE2A
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Sanger Sequencing
Quality control
- STR analysis
- TPOX, CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539
Cell culture
- Biosafety level
- EU: 2 US: 2
- Adherent/suspension
- Adherent
- Gender
- Female
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.
Storage
- Shipped at conditions
- Dry Ice
- Appropriate short-term storage conditions
- -196°C
- Appropriate long-term storage conditions
- -196°C
Notes
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.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
Rad6 also known as ubiquitin-conjugating enzyme E2 A (UBE2A) is an enzyme with a molecular weight of approximately 17 kDa. This enzyme plays an essential role in the ubiquitination process where it catalyzes the transfer of ubiquitin to substrate proteins. Rad6 is expressed in various tissues with significant activity observed in the testis and brain suggesting its involvement in cell division and development.
- Biological function summary
Rad6 functions as part of a larger protein complex involved in the DNA damage repair process. It collaborates with the RAD6 epistasis group which ensures the maintenance of genomic stability. Rad6 as a ubiquitin-conjugating enzyme interacts with E3 ubiquitin ligases facilitating the ubiquitination of target proteins that require degradation or regulation in response to DNA damage. This activity is vital for DNA repair mechanisms like post-replication repair (PRR) which protects cells from mutagenic events.
- Pathways
Rad6 integrates into critical biological processes including the ubiquitin-proteasome pathway and the DNA repair pathway. Within the ubiquitin-proteasome system Rad6 aids in tagging proteins for degradation thereby regulating protein levels and cellular functions. It connects to other proteins like E3 enzymes and RAD18 in the DNA repair pathway. Its activity in DNA repair ensures the appropriate response to DNA damage coordinating with other mechanistic proteins to maintain genomic stability.
- Associated diseases and disorders
Rad6 shows significant association with cancer and neurodegenerative conditions. Changes in Rad6 activity can lead to abnormal cell cycle progression contributing to oncogenesis particularly in breast and ovarian cancers. Rad6 interacts with proteins such as BRCA1 a tumor suppressor implicating its role in hereditary breast cancer. Additionally its dysfunction in the brain may relate to Alzheimer's disease highlighting its connection to neuroprotective pathways and protein degradation systems.
Product promise
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3 product images
Sanger Sequencing - Human UBE2A knockout HEK-293T cell line (ab266322)
Allele-1: 19 bp deletion in exon 1
Sanger Sequencing - Human UBE2A knockout HEK-293T cell line (ab266322)
Allele-2: 8 bp deletion in exon 1.
Sanger Sequencing - Human UBE2A knockout HEK-293T cell line (ab266322)
Sequencing chromatogram displaying sequence edit in exon 1
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