Human RB1CC1 (FIP200) knockout A549 cell line
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- NGS
Lab
Next Generation Sequencing - Human RB1CC1 (FIP200) knockout A549 cell line (AB277823)
4 bp deletion after Leu1377, 51 bp deletion downstream.
Complementary Products
Primary Antibodies
AB313621
Anti-FIP200 antibody [EPR26076-87] - BSA and Azide free
RabMAb|Recombinant
![Western blot - Anti-FIP200 antibody [EPR26076-87] - BSA and Azide free (AB313621)](https://content.abcam.com/products/images/fip200-antibody-epr26076-87-bsa-and-azide-free-ab313621--western-blot-img418384.jpg)
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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.
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.
What's included?
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.
- Do not allow the cell density to exceed 7x104 cells/cm2.
Culture medium
F-12K + 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
FIP200 is an important component of the ULK1 complex which is a critical player in the regulation of autophagy. Autophagy is a cellular degradation pathway that removes damaged organelles and proteins helping maintain cellular homeostasis. FIP200 interacts with other proteins such as ULK1 and ATG13 to facilitate the initiation of autophagy. Its role in this complex ensures proper cellular response to nutrient deprivation and stress conditions.
Pathways
The mechanistic functions of FIP200 integrate into the autophagy and mTOR signaling pathways. In the mTOR pathway FIP200 acts upstream interacting with proteins like mTOR kinase to coordinate cell growth and autophagy. This interaction contributes to maintaining cellular energy levels and adaptions to metabolic stress. FIP200's regulatory activities within these pathways impact various cellular functions aligning cell metabolism and survival mechanisms.
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'.
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