HERC5 KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 5 bp insertion in exon 1.
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Key facts
- Cell type
- A549
- Species or organism
- Human
- Tissue
- Lung
- Form
- Liquid
- Knockout validation
- Sanger Sequencing
- Mutation description
- Knockout achieved by using CRISPR/Cas9, Homozygous: 5 bp insertion in exon 1
Alternative names
CEB1, CEBP1, Cyclin-E-binding protein 1, E3 ISG15--protein ligase HERC5, HECT E3 ubiquitin ligase, HECT domain and RCC1-like domain-containing protein 5, HERC5_HUMAN, Hect domain and RLD 5
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HERC5 KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 5 bp insertion in exon 1.
Key facts
- Cell type
- A549
- Species or organism
- Human
- Tissue
- Lung
- Form
- Liquid
- Knockout validation
- Sanger Sequencing
- Mutation description
- Knockout achieved by using CRISPR/Cas9, Homozygous: 5 bp insertion in exon 1
- Disease
- Carcinoma
- Concentration
Properties
- Gene name
- HERC5
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Sanger Sequencing
- Zygosity
- Homozygous
Quality control
- STR analysis
- CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
- EU: 1 US: 1
- Adherent/suspension
- Adherent
- Gender
- Male
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.
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, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and 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
The HERC5 protein also known as HECT and RLD domain containing E3 ubiquitin protein ligase 5 functions mechanically as an E3 ubiquitin ligase involved in the post-translational modification process called ISGylation. It facilitates the attachment of ubiquitin-like protein ISG15 to target proteins. HERC5 has a molecular mass of approximately 112 kDa. Expression of HERC5 is mainly in human cells especially in high quantities in the spleen lung and kidney. The expression is often elevated by type I interferons indicating a role in immune response.
- Biological function summary
The HERC5 protein contributes to innate immune response by modifying proteins involved in antiviral defense. It adds ISG15 to newly synthesized proteins marking them for rapid degradation or altering their activities. HERC5 operates as a solitary protein rather than forming part of a larger complex. Through its ISGylation activity HERC5 plays a significant role in enhancing the antiviral state of cells inhibiting viral replication and modulating cellular processes during stress responses.
- Pathways
HERC5 actively integrates into the antiviral defense mechanism and ubiquitin-proteasome pathway. It associates with pathways related to protein modification and immune signaling significantly affecting how cells respond to viral infections. Proteins like ISG15 and ubiquitin-conjugating enzymes interact with HERC5 in these pathways highlighting its importance in maintaining cellular homeostasis and protecting against pathogens.
- Associated diseases and disorders
The HERC5 protein shows connections with viral infections and certain inflammatory conditions. Its heightened activity associates with viral diseases like influenza and hepatitis where enhanced ISGylation occurs as a defensive strategy against viral proliferation. Misregulation of ISGylation through HERC5 also relates to neuroinflammatory disorders suggesting that targeting this pathway may offer therapeutic potential. Researchers continue exploring the interaction of HERC5 with proteins linked to these conditions to better understand its impact on disease progression and potential interventions.
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1 product image
Sanger Sequencing - Human HERC5 knockout A549 cell line (ab267201)
Homozygous: 5 bp insertion in exon1
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