NOTCH1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 14 bp deletion in exon 6 and 1 bp deletion in exon 6 and 1 bp insertion in exon 6.
Images
Key facts
- Cell type
- HeLa
- Species or organism
- Human
- Tissue
- Cervix
- Form
- Liquid
- Knockout validation
- Sanger Sequencing, Western blot
- Mutation description
- Knockout achieved by using CRISPR/Cas9, 14 bp deletion in exon 6 and 1 bp deletion in exon 6 and 1 bp insertion in exon 6
Recommended products
NOTCH1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 14 bp deletion in exon 6 and 1 bp deletion in exon 6 and 1 bp insertion in exon 6.
Key facts
- Cell type
- HeLa
- Species or organism
- Human
- Tissue
- Cervix
- Form
- Liquid
- Knockout validation
- Sanger Sequencing, Western blot
- Mutation description
- Knockout achieved by using CRISPR/Cas9, 14 bp deletion in exon 6 and 1 bp deletion in exon 6 and 1 bp insertion in exon 6
- Disease
- Adenocarcinoma
- Concentration
Properties
- Gene name
- NOTCH1
- 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 HeLa cell line (ab255448). 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
Notch1 also known as Notch-1 is a transmembrane receptor involved in cell fate decisions. It belongs to the Notch protein family and has a molecular weight of around 270 kDa. Notch1 is expressed widely particularly in tissues such as blood cells and neuronal tissue. The receptor consists of extracellular EGF-like repeats a transmembrane domain and an intracellular domain that translocates to the nucleus upon activation. Notch1 plays a critical role in intercellular communication through ligand engagement which then triggers a series of proteolytic cleavages that release the Notch intracellular domain (NICD) for nuclear translocation.
- Biological function summary
Notch1 is essential in various cellular processes including differentiation proliferation and apoptosis. It functions as part of a complex signaling pathway that regulates these processes. Notch1 interacts with other elements like Jagged and Delta/Serrate/LAG-2 (DSL) family ligands to control gene expression patterns that determine cell lineage outcomes. This interaction affects the development of many systems such as the immune and nervous systems. Consequently Notch1 significantly influences the formation of organs and the maintenance of stem cell populations.
- Pathways
Notch1 plays a significant role in both the Notch signaling and the Wnt signaling pathways. In the Notch signaling pathway Notch1 upon ligand binding partners with CSL (CBF1/RBP-Jκ in mammals) and Mastermind-like proteins for transcriptional regulation. This pathway interlinks with the Wnt pathway that involves proteins like β-catenin affecting the regulation of gene transcription. The interplay between Notch1 and these pathways is fundamental in determining outcomes in cell proliferation and differentiation emphasizing the interconnected nature of signaling networks.
- Associated diseases and disorders
Notch1 is associated with T-cell acute lymphoblastic leukemia and breast cancer. Altered Notch1 signaling often through gain-of-function mutations can drive oncogenesis by disrupting normal cell differentiation and promoting uncontrolled proliferation. In T-cell acute lymphoblastic leukemia aberrant activation of Notch1 leads to increased expression of target genes working closely with related proteins like c-Myc. In breast cancer dysregulation of Notch1 signaling may facilitate tumor growth and metastasis indicating its role as a therapeutic target.
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3 product images
Sanger Sequencing - Human NOTCH1 knockout HeLa cell line (ab265651)
Allele-1: 1 bp insertion in exon 6.
Sanger Sequencing - Human NOTCH1 knockout HeLa cell line (ab265651)
Allele-2: 14 bp deletion in exon 6.
Sanger Sequencing - Human NOTCH1 knockout HeLa cell line (ab265651)
Allele-3: 1 bp deletion in exon 6.
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