#151828

HEK293-A mRFP-ATG9 Cell Line

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Cat. #151828

HEK293-A mRFP-ATG9 Cell Line

Cat. #: 151828

Sub-type: Continuous

Unit size: 1x10^6 cells / vial

Availability: 3-5 days

Organism: Human

Tissue: Embryo

Model: Reporter

£575.00

This fee is applicable only for non-profit organisations. If you are a for-profit organisation or a researcher working on commercially-sponsored academic research, you will need to contact our licensing team for a commercial use license.

Contributor

Inventor: Sharon Tooze

Institute: Cancer Research UK, London Research Institute: Lincoln's Inn Fields

Tool Details
Target Details
Applications
Handling
References

Tool Details

*FOR RESEARCH USE ONLY

  • Name: HEK293-A mRFP-ATG9 Cell Line
  • Research fields: Apoptosis and autophagy;Cancer;Cell biology;Cell signaling and signal transduction;Drug development;Metabolism
  • Tool sub type: Continuous
  • Parental cell: HEK293-A
  • Organism: Human
  • Tissue: Embryo
  • Model: Reporter
  • Conditional: Yes
  • 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.
  • 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.
  • Biosafety level: 1
  • Additional notes: HEK293 cells were obtained under licence from AdVec
  • Recommended controls: HEK293-A parental line
  • Cellosaurus id: CVCL_W349

Target Details

  • Target: ATG9A

Applications

  • Application notes: HEK293 cells were obtained under licence from AdVec

Handling

  • Format: Frozen
  • Growth medium: DMEM + 10% FCS + 2mM Glutamine + 400ug/ml geneticin (G418) for maintainance.
  • Unit size: 1x10^6 cells / vial
  • Shipping conditions: Dry ice
  • Storage conditions: Liquid Nitrogen
  • Mycoplasma free: Yes

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.