ATRIP KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 13 bp deletion in exon 4 and 1 bp insertion in exon 4 and 7 bp deletion in exon 4.
Images
Key facts
- Cell type
- HEK-293T
- Species or organism
- Human
- Tissue
- Kidney
- Form
- Liquid
- Knockout validation
- Sanger Sequencing, Western blot
- Mutation description
- Knockout achieved by using CRISPR/Cas9, 13 bp deletion in exon 4 and 1 bp insertion in exon 4 and 7 bp deletion in exon 4
Recommended products
ATRIP KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 13 bp deletion in exon 4 and 1 bp insertion in exon 4 and 7 bp deletion in exon 4.
Key facts
- Cell type
- HEK-293T
- Species or organism
- Human
- Tissue
- Kidney
- Form
- Liquid
- Knockout validation
- Sanger Sequencing, Western blot
- Mutation description
- Knockout achieved by using CRISPR/Cas9, 13 bp deletion in exon 4 and 1 bp insertion in exon 4 and 7 bp deletion in exon 4
- Concentration
Properties
- Gene name
- ATRIP
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Sanger Sequencing, Western blot
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, 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
ATRIP also known as ATRIP protein serves as an essential component within the ATR-ATRIP complex which is vital in the DNA damage response. ATRIP is a protein that weighs approximately 86 kDa and is most commonly expressed in the nucleus of cells. It functions primarily by binding to ATR (Ataxia Telangiectasia and Rad3-related protein) which enables the ATR to localize to sites of DNA damage and replication stress. This enables the recognition and repair of damaged DNA preventing genomic instability.
- Biological function summary
ATRIP plays roles in maintaining genome integrity. It forms a stable complex with ATR which is important for the activation of ATR kinase activity. ATRIP directly interacts with single-stranded DNA coated by replication protein A (RPA) to perform its functions. By facilitating ATR recruitment to DNA damage sites ATRIP aids in the initiation of DNA repair mechanisms which are critical for normal cellular function and survival.
- Pathways
ATRIP's activities are an integral part of the DNA damage signaling and repair pathways. ATR-ATRIP complex is central to the DNA replication checkpoint pathway where it maintains replication fork stability and controls cell cycle progression. Additionally ATRIP is involved in pathways alongside proteins like CHK1 which it helps to activate in response to DNA damage ensuring proper cell cycle arrest and repair processes.
- Associated diseases and disorders
ATRIP is implicated in certain cancers and genomic instability syndromes. Deficient ATRIP function can correlate with a higher predisposition to cancers due to defective DNA damage response and repair. Further the ATR-ATRIP interaction can influence the onset and progression of Seckel syndrome where defects in ATRIP or its partner ATR might lead to developmental abnormalities. The role of ATRIP in these diseases highlights the importance of maintaining genomic stability through effective DNA repair mechanisms.
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3 product images
Sanger Sequencing - Human ATRIP knockout HEK-293T cell line (ab267261)
Allele-2: 7 bp deletion in exon 4.
Sanger Sequencing - Human ATRIP knockout HEK-293T cell line (ab267261)
Allele-1: 13 bp deletion in exon4
Sanger Sequencing - Human ATRIP knockout HEK-293T cell line (ab267261)
Allele-3: 1 bp insertion in exon 4.
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