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AB265140

Human UBE3B knockout HeLa cell line

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UBE3B KO cell line available to order. KO validated by. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 2 bp deletion in exon 3.

View Alternative Names

AI449831, AU020130, FLJ45294, MGC131858, MGC78388, Ubiquitin protein ligase E3B

2 Images
Sanger Sequencing - Human UBE3B knockout HeLa cell line (AB265140)
  • Sanger seq

Unknown

Sanger Sequencing - Human UBE3B knockout HeLa cell line (AB265140)
Sanger Sequencing - Human UBE3B knockout HeLa cell line (AB265140)
  • Sanger seq

Unknown

Sanger Sequencing - Human UBE3B knockout HeLa cell line (AB265140)

Allele-1 : 2 bp deletion in exon 3.

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Form

Liquid

form

Knockout validation

Sanger Sequencing

Mutation description

Knockout achieved by using CRISPR/Cas9, 2 bp deletion in exon 3

Disease

Adenocarcinoma

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 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?

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Properties and storage information

Gene name
UBE3B
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing
Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C

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.

UBE3B also known as ubiquitin-protein ligase E3B or hRPF1 is an enzyme that catalyzes the transfer of ubiquitin from an E2 ubiquitin-conjugating enzyme to a substrate protein tagging it for degradation by the proteasome. It has a molecular weight of approximately 120 kDa. UBE3B is broadly expressed across various tissues including brain heart and muscles. The widespread expression pattern emphasizes its importance in protein homeostasis across different cell types.
Biological function summary

UBE3B functions as an important regulator in cellular processes by tagging damaged or unneeded proteins with ubiquitin directing them for degradation. This protein is not part of any known large macromolecular complex but works closely with other enzymes in the ubiquitination cascade especially E2 enzymes. Through this activity UBE3B helps control the quality of proteins within the cell aiding in the removal of proteins that could potentially be harmful.

Pathways

UBE3B plays a significant role in the ubiquitin-proteasome pathway which is pivotal for protein turnover and cellular regulation. Additionally UBE3B interacts with pathways involved in synaptic function highlighting its possible connection to neurological processes. It works alongside other ligases and deubiquitinating enzymes facilitating the maintenance of cellular protein levels and functions that are necessary for proper signaling and response in cells.

Mutations in UBE3B have been linked to neurological disorders including Kaufman oculocerebrofacial syndrome. This condition highlights the protein's involvement in brain development and function. UBE3B's ubiquitination activity connects it with proteins like SHANK3 which is also implicated in neurodevelopmental disorders. Therefore disturbances in UBE3B's function can have significant consequences for neural integrity and development.

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 protocols

Product promise

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