Human UBE2A knockout HEK-293T cell line
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- Sanger seq
Unknown
Sanger Sequencing - Human UBE2A knockout HEK-293T cell line (AB266322)
Allele-1 : 19 bp deletion in exon 1
- Sanger seq
Lab
Sanger Sequencing - Human UBE2A knockout HEK-293T cell line (AB266322)
Sequencing chromatogram displaying sequence edit in exon 1
- Sanger seq
Unknown
Sanger Sequencing - Human UBE2A knockout HEK-293T cell line (AB266322)
Allele-2 : 8 bp deletion in exon 1.
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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
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
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.
Quality control
STR analysis
TPOX, CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Target data
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