ING2 KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 11 bp deletion in exon 1.
Images
Key facts
- Cell type
- HEK-293T
- Species or organism
- Human
- Tissue
- Kidney
- Form
- Liquid
- Knockout validation
- Sanger Sequencing
- Mutation description
- Knockout achieved by using CRISPR/Cas9, Homozygous: 11 bp deletion in exon 1
Recommended products
ING2 KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 11 bp deletion in exon 1.
Key facts
- Cell type
- HEK-293T
- Species or organism
- Human
- Tissue
- Kidney
- Form
- Liquid
- Knockout validation
- Sanger Sequencing
- Mutation description
- Knockout achieved by using CRISPR/Cas9, Homozygous: 11 bp deletion in exon 1
- Concentration
Properties
- Gene name
- ING2
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Sanger Sequencing
Cell culture
- Biosafety level
- EU: 2 US: 2
- Viability
- ~ 80%
- 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
Although we aim to provide customers with a homozygous clone, feasibility will be dependent on the biology of the protein. Should only heterozygous edits be achieved, you will be notified of the outcome and be asked to confirm whether the cell line is acceptable. All clones will be accompanied with DNA sequencing data, and the mutation description.
Recommended control: Human wild-type HEK293T cell line (ab255449). 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.
Cryopreservation cell medium: Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Culture medium: DMEM (High Glucose) + 10% FBS
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 2x10E4 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 2x10E4 cells/cm2 is recommended.
• Cells should be passaged when they have achieved 80-90% confluence.
• Do not allow the cell density to exceed 7x10E4 cells/cm2.
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
ING2 also known as Inhibitor of Growth Protein 2 is a nuclear protein with a molecular weight of approximately 33 kDa. It expresses highly in tissues such as the prostate placenta and lung. Nuclear localization signals and PHD-finger motifs characterize its structure allowing it to bind to chromatin and influence gene expression. As a member of the ING family ING2 plays a role in regulating various cellular processes through its presence in different cellular compartments.
- Biological function summary
ING2 functions as a tumor suppressor by controlling the transcription of genes involved in cell cycle regulation and apoptosis. It interacts with histone acetyltransferase (HAT) and histone deacetylase (HDAC) complexes contributing to chromatin remodeling. As part of the Sin3A-HDAC complex ING2 has the ability to repress transcription influencing the expression of genes important for maintaining cellular homeostasis and preventing oncogenesis.
- Pathways
The ING2 protein actively participates in the p53 signaling pathway as well as the MAPK signaling pathway. Its interaction with p53 leads to the enhancement of p53-mediated transcriptional activation which promotes cell cycle arrest and apoptosis. ING2 also interacts with proteins like TP53 and EP300 modulating the cellular responses to stress signals and contributing to cell survival and repair mechanisms.
- Associated diseases and disorders
The involvement of ING2 has been linked to cancer and neurodegenerative disorders. In cancer downregulation or mutation of ING2 disrupts the p53 pathway leading to impaired tumor suppression. ING2's interaction with TP53 and its tumor suppressor role highlights its importance in cancer biology. Furthermore in neurodegenerative disorders ING2's dysregulation can influence neuronal survival pathways potentially affecting proteins like p53 and EP300 therefore altering neuronal function and contributing to disease progression.
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1 product image
Sanger Sequencing - Human ING2 knockout HEK-293T cell line (ab267317)
Homozygous: 11 bp deletion in exon1
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