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Contributor Information

  • Name T.R. Santhosh Kumar
  • Institute Rajiv Gandhi Centre for Biotechnology
  • Primary citation Srinivas KP, et.al. PMID:26992219

Tool Details

  • Tool name: SiHa/EGFP_Cytochrome c
  • Tool type: Cell Lines
  • Parental cell line: SiHa
  • Organism: Human
  • Tissue: Cervix
  • Gender: Female
  • Cancer type: Gynaecologic cancer
  • Disease: Cancer
  • Morphology: Epithelial
  • Growth properties: Adherent
  • Model: Transgenic
  • CRISPR: No
  • Conditional: No
  • Description: The product is a human cervical carcinoma cell line SiHa, transfected to stably express the protein cytochrome c (cyt-c) tagged with EGFP. The EGFP-tag enables easy identification of the cyt-c release in live cells upon apoptotic induction. The cell line can be utilized for studying the apoptosis process and to screen of potential candidate drugs that induce apoptosis. Stably expressing single cell clones with homogeneous expression of Cytochrome c_EGFP were isolated for high resolution imaging. The expression and functionality of exogenous Cytochrome c_EGFP was confirmed through fluorescence microscopy by its release upon apoptotic stimulus.
  • Research area: Cancer; Disease (not cancer); Metabolism

  • For Research Use Only

Target Details

Application Details

Handling

  • Growth medium: DMEM with 2mM L-Glutamine and 10% FBS (Heat inactivated), or EMEM with 2mM L-Glutamine and 10% Fetal Bovine Serum.
  • Temperature: 37C
  • Atmosphere: 5% CO2
  • Storage medium: Growth medium + 10% DMSO
  • Storage conditions: Liquid Nitrogen
  • Shipping conditions: Dry Ice
  • Initial handling information: Resuscitation: Rapidly thaw the frozen ampoule in a water bath at 37?‚°C for 1-2 minutes. Transfer the contents to a centrifuge tube and slowly add 5-10ml of pre-warmed growth media. Remove a sample for counting. Centrifuge at 100-150g for 2-3 minutes to pellet cells (a centrifugation step to remove the cryoprotectant is essential. Seed at the recommend density, e.g. greater than 10,000 cells per cm2.
  • Cultured in antibiotics?: No
  • Subculture routine: Subculture non-confluent cultures using trypsin/EDTA and 1:3 to 1:4, i.e. seeding at 1-4 x10,000 cells/cm?‹Â› .

Documentation

References

  •   Srinivas KP, et.al. PMID:26992219