NRAS KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 1 and 2 bp deletion in exon 1 and 5 bp deletion in exon 1.
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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, 1 bp insertion in exon 1 and 2 bp deletion in exon 1 and 5 bp deletion in exon 1
Alternative names
ALPS4, AV095280, GTPase NRas, H-Ras-1, N ras, N ras protein part 4, NRAS1, NS6, Neuroblastoma RAS viral (v ras) oncogene homolog, OTTHUMP00000013879, OTTMUSP00000023521, RASN_HUMAN, Transforming protein N-Ras, v ras neuroblastoma RAS viral oncogene homolog
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NRAS KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 1 and 2 bp deletion in exon 1 and 5 bp deletion in exon 1.
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, 1 bp insertion in exon 1 and 2 bp deletion in exon 1 and 5 bp deletion in exon 1
- Disease
- Adenocarcinoma
- Concentration
Properties
- Gene name
- NRAS
- 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, 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 NRAS protein also known as neuroblastoma RAS viral oncogene homolog has a molecular weight of approximately 24 kDa. It functions mechanically as a GTPase cycling between an inactive GDP-bound state and an active GTP-bound state. The NRAS gene which encodes this protein shows expression in various tissues including the thymus leukocytes and other cells of the hematopoietic lineage. Activation of RAS involves the conversion to the GTP-bound form which enables NRAS to transmit signals within cells.
- Biological function summary
NRAS plays an important role in cell proliferation differentiation and survival. It does not function alone but is part of the larger RAS protein family including HRAS and KRAS with whom it shares similar roles and sequence homology. NRAS interacts with various growth factor receptors and mediates signals to downstream effectors that influence cellular processes. Mutations in the NRAS gene can affect these biological processes leading to altered cell behavior.
- Pathways
NRAS has significant roles in the MAPK/ERK pathway and the PI3K/AKT pathway. These pathways regulate fundamental cellular functions including growth and survival. NRAS interacts closely with other proteins such as RAF kinases and PI3K to propagate signals from activated receptors at the cell membrane to the nucleus. Altered NRAS activity can impact these pathways affecting cellular responses to external stimuli.
- Associated diseases and disorders
NRAS mutations are linked to certain cancers including melanoma and acute myeloid leukemia (AML). Mutant forms of NRAS result in constitutively activated RAS contributing to uncontrolled cell growth and survival. In melanoma NRAS mutations frequently co-occur with alterations in other proteins such as BRAF another member of the RAS/RAF/MEK/ERK pathway. Understanding NRAS's involvement in these conditions is critical for developing targeted therapeutic strategies.
Product promise
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3 product images
Sanger Sequencing - Human NRAS knockout HeLa cell line (ab267224)
Allele-3: 5 bp deletion in exon 1.
Sanger Sequencing - Human NRAS knockout HeLa cell line (ab267224)
Allele-1: 2 bp deletion in exon1
Sanger Sequencing - Human NRAS knockout HeLa cell line (ab267224)
Allele-2: 1 bp insertion in exon 1.
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