RB1CC1 KO cell line available to order. KO validated by Next Generation Sequencing. Free of charge wild type control provided.
FAK family kinase-interacting protein of 200 kDa, FIP200, RB1-inducible coiled-coil protein 1, RB1CC1, RBCC1_HUMAN
RB1CC1 KO cell line available to order. KO validated by Next Generation Sequencing. Free of charge wild type control provided.
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.
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 A549 cell line (ab288558). 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.
FIP200 also known as RB1CC1 is a large protein with a mass of approximately 200 kDa. It functions in several cellular processes acting as a scaffold protein that assists in the assembly of different protein complexes. FIP200 is widely expressed in human tissues including the brain heart and skeletal muscle. This suggests it has important roles in diverse biological functions within the body.
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.
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.
FIP200's dysfunction has connections to cancer and neurodegenerative diseases. In cancer alterations in FIP200 expression can affect cell proliferation and tumorigenesis possibly through associations with proteins like p53. In neurodegenerative diseases improper autophagy due to malfunctioning FIP200 can lead to the accumulation of damaged proteins and cellular stress forging a link with disorders such as Alzheimer's. Understanding FIP200’s role in these conditions aids the exploration of therapeutic targets for disease treatment.
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4 bp deletion after Leu1377, 51 bp deletion downstream.
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