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AB267178

Human USP18 (UBP43) knockout A549 cell line

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USP18 KO cell line available to order. KO validated. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 97 bp deletion in exon 4. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
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Sanger Sequencing - Human USP18 (UBP43) knockout A549 cell line (AB267178)
  • Sanger seq

Unknown

Sanger Sequencing - Human USP18 (UBP43) knockout A549 cell line (AB267178)

Homozygous : 97 bp deletion in exon4

Key facts

Cell type

A549

Species or organism

Human

Tissue

Lung

Form

Liquid

form

Knockout validation

Sanger Sequencing

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: 97 bp deletion in exon 4

Disease

Carcinoma

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?

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Properties and storage information

Gene name
USP18
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing
Zygosity
Homozygous
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.
  • Do not allow the cell density to exceed 7x104 cells/cm2.
Culture medium

F-12K + 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.

UBP43 also known as USP18 is a deubiquitinating enzyme that is 43 kilodaltons in mass. This protein removes ubiquitin from ubiquitinated proteins which regulates their degradation and stability. UBP43 is expressed widely in the body with significant levels in the immune and nervous systems. Its alternate name USP18 reflects its inclusion in the ubiquitin-specific protease family highlighting its specific function in modulating protein ubiquitination.
Biological function summary

UBP43 regulates type I interferon signaling and affects immune response. As a negative feedback regulator it inhibits the interferon signaling pathway preventing unnecessary immune reactions. UBP43 does not form a part of a larger protein complex but acts independently to maintain cellular homeostasis. Its activity impacts the function of other proteins involved in response to viral infections.

Pathways

UBP43 plays an important role in the JAK-STAT signaling pathway interacting with proteins involved in cytokine signaling. It influences the pathway by inhibiting the action of interferon-stimulated genes therefore modulating the immune response. UBP43's interaction with ISG15 a ubiquitin-like modifier is also a critical aspect that affects other cellular processes like protein modification and response to inflammatory stimuli.

UBP43 is closely linked to autoimmune diseases and viral infections. Altered UBP43 activity can lead to improper immune responses contributing to conditions such as systemic lupus erythematosus. It also plays a role in the body's defense against viral infections where its dysregulation may affect the efficiency of antiviral responses. The interplay between UBP43 and ISG15 is of particular interest as changes in their interaction could provide insights into the development or progression of these disorders.

Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

Cell culture

Biosafety level

EU: 1 US: 1

Adherent/suspension

Adherent

Gender

Male

Product protocols

Product promise

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