Human NF1 knockout MCF7 cell line
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NF1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. To order both knockout and wild-type control cells: select 2 x 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.
View 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
- WB
Lab
Western blot - Human NF1 knockout MCF7 cell line (AB286416)
Anti-NF1 antibody [EPR22989-68] (ab238142) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab238142 was shown to bind specifically to NF1. A band was observed at 180-270 kDa in wild-type MCF7 cell lysates with no signal observed at this size in NF1 knockout cell line. To generate this image, wild-type and NF1 knockout MCF7 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
All lanes:
Western blot - Anti-Neurofibromin antibody [EPR22989-68] (<a href='/en-us/products/primary-antibodies/neurofibromin-antibody-epr22989-68-ab238142'>ab238142</a>) at 1/1000 dilution
Lanes 1 - 4:
Western blot at 20 µg
Lane 2:
Western blot - Human NF1 knockout MCF7 cell line (ab286416) at 20 µg
Secondary
All lanes:
Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Predicted band size: 319 kDa
false
- NGS
Lab
Next Generation Sequencing - Human NF1 knockout MCF7 cell line (AB286416)
151 bp deletion after the Ile108 of the WT protein
Product details
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 MCF7 cell line (ab288560). 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.
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
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
- Slow to trypsinise.
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 5-7x104 cells/cm2 is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
Culture medium
MEM + 10% FBS + 0.01 mg/ml bovine insulin
Cryopreservation medium
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
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.
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Female
Target data
Complementary Products
Primary Antibodies
AB238142
Anti-Neurofibromin antibody [EPR22989-68]
RabMAb|Recombinant|KO Validated
![Western blot - Anti-Neurofibromin antibody [EPR22989-68] (AB238142)](https://content.abcam.com/products/images/neurofibromin-antibody-epr22989-68-ab238142--western-blot-img419782.jpg)
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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