Human NANOG knockout HCT116 cell line
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- NGS
Lab
Next Generation Sequencing - Human NANOG knockout HCT116 cell line (AB289096)
106 bp deletion after Cys 12 (allele 1); 8 bp deletion after Cys 12 (allele 2) of WT protein
Complementary Products
Primary Antibodies
AB109250
Anti-Nanog antibody [EPR2027(2)]
RabMAb|Advanced Validation|Recombinant
![ChIP-sequencing - Anti-Nanog antibody [EPR2027(2)] (AB109250)](https://content.abcam.com/products/images/nanog-antibody-epr20272-ab109250--chip-sequencing-img402752.jpg)
- 4
- 11
Reactivity data
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 HCT116 cell line (ab288559). 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
- 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
McCoY5a + 10% FBS
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
Nanog plays a central role in stem cell biology operating as part of a complex regulatory network. Nanog interacts with other important transcription factors like Oct4 and Sox2 forming a core pluripotency network. This network suppresses the differentiation of stem cells by regulating the expression of genes involved in cell fate decisions. By modulating the expression of different pathways Nanog ensures the maintenance of an undifferentiated state in stem cells.
Pathways
Nanog is deeply embedded in the Wnt/β-Catenin and TGF-β signaling pathways important for stem cell maintenance and differentiation. In the Wnt/β-Catenin pathway Nanog works alongside proteins like β-catenin to drive the expression of genes that promote self-renewal. Meanwhile in the TGF-β pathway Nanog acts with proteins such as Smad2/3 to balance pluripotency and differentiation signals. These pathways critically support the complex network that sustains stem cell identity and function.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
Target data
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com