The HEK293-A mRFP-ATG9 cell line is a stable cell line expressing mRFP-tagged ATG9 (human) in background of HEK293A cell line. It can be used to monitor delivery of membrane from the endosomes and Golgi to the forming autophagosome using live-cell microscopy and biochemical approaches.
| Inventor | Institute |
|---|---|
| Sharon Tooze | Cancer Research UK, London Research Institute: Lincoln's Inn Fields |
| SKU: | 151828 |
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| Product description: | The HEK293-A mRFP-ATG9 cell line is a stable cell line expressing mRFP-tagged ATG9 (human) in background of HEK293A cell line. It can be used to monitor delivery of membrane from the endosomes and Golgi to the forming autophagosome using live-cell microscopy and biochemical approaches. |
| Conditional: | Yes |
| Production details: | pEGFP-mRFP-Atg9 plasmid was transfected into HEK293A cells. The plasmid was generated by excision of the EGFP coding sequence from pEGFP-C1 and insertion of the mRFP-ATG9 cassette. The cells were selected with 800ug/ml geneticin (G418) followed by single cell clonal dilution. Clones were screened by confocal microscopy for low-level expression in which mRFP- Atg9 distribution resembled that of endogenous mAtg9 and by western blot. Clone 1F10 was selected as the best based on these parameters. |
| Additional notes: | HEK293 cells were obtained under licence from AdVec |
| Cellosaurus ID: | CVCL_W349 |
| Parental cell line: | HEK293-A |
| Cat. #: | 151828 |
|---|---|
| Provenance and Ownership: | The HEK293-A mRFP-ATG9 Cell Line is owned by Cancer Research UK. CancerTools supplies the HEK293-A mRFP-ATG9 Cell Line under licence from Cancer Research UK and is the authorised source for its distribution. Sourcing HEK293-A mRFP-ATG9 Cell Line directly from the IP owner ensures authenticity, provenance and compliance with all applicable licensing and research use requirements. |
| Tool sub type: | Continuous |
| Unit size: | 1×10^6 cells / vial |
| Research Fields: | Apoptosis and autophagy; Cancer; Cell biology; Cell signaling and signal transduction; Drug development; Metabolism |
| Organism: | Human |
| Tissue: | Embryo |
| Model: | Reporter |
| Format: | Frozen |
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| Storage conditions: | Liquid Nitrogen |
| Shipping conditions: | Dry ice |
| Growth medium: | DMEM + 10% FCS + 2mM Glutamine + 400ug/ml geneticin (G418) for maintainance. |
| Recommended controls: | HEK293-A parental line |
| Mycoplasma free: | Yes |
| Biosafety level: | 1 |
| References: |
Orsi et al. 2012. Mol Biol Cell. 23(10):1860-73. PMID: 22456507. Dynamic and transient interactions of Atg9 with autophagosomes, but not membrane integration, are required for autophagy. Young et al. 2006. J Cell Sci. 119(Pt 18):3888-900. PMID: 16940348. Starvation and ULK1-dependent cycling of mammalian Atg9 between the TGN and endosomes. |
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