DPH2 KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 20 bp deletion in exon 2 and 7 bp deletion in exon 2.
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Key facts
- Cell type
- HeLa
- Species or organism
- Human
- Tissue
- Cervix
- Form
- Liquid
- Knockout validation
- Sanger Sequencing
- Mutation description
- Knockout achieved by using CRISPR/Cas9, 20 bp deletion in exon 2 and 7 bp deletion in exon 2
Alternative names
DPH 2 homolog, DPH2 homolog S. cerevisiae, DPH2 like 2 S. cerevisiae, DPH2-like 2, DPH2L2, DPH2_HUMAN, Diphthamide biosynthesis like protein 2, Diphthamide biosynthesis protein 2, Diphthamide biosynthesis protein 2 homolog-like 2, Diptheria toxin resistance protein required for diphthamide biosynthesis like 2, Diptheria toxin resistance protein required for diphthamide biosynthesis like 2 S. cerevisiae, HsDph2, OTTHUMP00000010007, Protein DPH2 homolog
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DPH2 KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 20 bp deletion in exon 2 and 7 bp deletion in exon 2.
Key facts
- Cell type
- HeLa
- Species or organism
- Human
- Tissue
- Cervix
- Form
- Liquid
- Knockout validation
- Sanger Sequencing
- Mutation description
- Knockout achieved by using CRISPR/Cas9, 20 bp deletion in exon 2 and 7 bp deletion in exon 2
- Disease
- Adenocarcinoma
- Concentration
Properties
- Gene name
- DPH2
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Sanger Sequencing
Quality control
- STR analysis
- CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
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
DPH2 also known as diphthamide biosynthesis protein 2 functions mechanically in catalyzing the first step of diphthamide biosynthesis. The protein incorporates a unique post-translational modification into histidine residues on elongation factor 2 (EEF2). DPH2 weighs approximately 53 kDa and is expressed in various tissues with high levels seen in the liver and testis. Its structural genes reside on human chromosome 17.
- Biological function summary
Diphthamide biosynthesis is important to protein synthesis and cellular resilience against bacterial toxins like diphtheria toxin. DPH2 operates within a multiprotein complex including DPH1 and DPH3 necessary for EEF2 modification. Proper function of DPH2 ensures that EEF2 maintains its role in translational fidelity which is indispensable for normal cellular processes.
- Pathways
Diphthamide biosynthesis connects to the protein synthesis and modification pathway. DPH2's activities integrate seamlessly with the translational machinery specifically influencing EEF2's function in the ribosome. In addition DPH2 works with DPH1 DPH3 and other proteins in the biosynthesis pathway to fulfill its role efficiently supporting cellular defense mechanisms against translational errors induced by toxins.
- Associated diseases and disorders
Defects in DPH2 relate to cancer and neurological conditions. Loss of function mutations in DPH2 can affect cellular processes potentially contributing to tumorigenesis where DPH1 is another connected protein of interest. Genetic alterations in DPH2 can also interfere with neural cell proliferation and survival linking to certain neurodegenerative disorders and emphasizing its role in maintaining cell health.
Product promise
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2 product images
Sanger Sequencing - Human DPH2 knockout HeLa cell line (ab265657)
Allele-2: 7 bp deletion in exon 2.
Sanger Sequencing - Human DPH2 knockout HeLa cell line (ab265657)
Allele-1: 20 bp deletion in exon 2.
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