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NF1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon 1.

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Images

Western blot - Human NF1 (Neurofibromin) knockout HeLa cell line (AB264725), expandable thumbnail
  • Sanger Sequencing - Human NF1 (Neurofibromin) knockout HeLa cell line (AB264725), expandable thumbnail

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, Homozygous: 1 bp deletion in exon 1

Alternative names

DKFZp686J1293, FLJ21220, NF, NF1_HUMAN, NFNS, Neurofibromatosis Noonan syndrome, Neurofibromatosis-related protein NF-1, Neurofibromin 1, Neurofibromin truncated, Type 1 Neurofibromatosis, VRNF, WATS, WSS, Watson disease related protein WSS, Watson syndrome, neurofibromatosis type I, von Recklinghausen disease neurofibromin, von Recklinghausen disease related protein VRNF

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NF1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon 1.

Key facts

Cell type
HeLa
Form
Liquid
Mutation description
Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon 1
Disease
Adenocarcinoma
Concentration
Loading...

Properties

Gene name
NF1
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Zygosity
Homozygous

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

Recommended control: Human wild-type HeLa cell line (ab255928). 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.

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

Neurofibromin is a protein that functions as a Ras GTPase-activating protein acting to promote the hydrolysis of Ras-bound GTP to GDP which inactivates Ras signaling. This regulatory function involves the suppression of excessive cell proliferation and differentiation. Alternate names for neurofibromin include NF1 or neurofibromin 1. The mass of neurofibromin is approximately 327 kDa. Neurofibromin is expressed widely across various tissues with notable expression in the nervous system and the brain.

Biological function summary

Neurofibromin regulates pathways involved in cell growth and development. It does not form part of a large multiprotein complex but interacts with microtubules and other signaling proteins to carry out its regulatory functions. Neurofibromin primarily affects pathways that manage cell cycle progression and it plays a critical role in inhibiting uncontrolled cell growth ensuring proper cellular homeostasis.

Pathways

Neurofibromin fits into the Ras-MAPK pathway where it controls the activity of Ras an important regulatory protein involved in signal transduction related to cell division and differentiation. Neurofibromin's action on Ras directly impacts the activation of the MAPK pathway which is important in mediating cellular responses to growth signals. This protein also interacts functionally with proteins like SOS1 another regulator of Ras emphasizing its role in fine-tuning signal cascades that ensure balanced cellular function.

Associated diseases and disorders

Neurofibromin's dysregulation is most commonly associated with neurofibromatosis type 1 a genetic disorder characterized by the development of benign tumors in the nervous system. The loss of neurofibromin function leads to unregulated Ras activity resulting in tumorigenesis. Additionally neurofibromin is implicated in certain sporadic cancers due to its role in controlling cell proliferation. In neurofibromatosis type 1 interactions between neurofibromin and proteins such as p53 a known tumor suppressor highlight its significance in disease pathways and tumor suppression mechanisms.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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