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Contributor Information

  • Name Jonathon Pines
  • Institute University of Cambridge

Tool Details

  • Tool name: Venus-Mad2 Reporter Cell Line [RPE1 Venus-Mad2/+ KI clone #1]
  • Alternate names: MAD2L1
  • Tool type: Cell Lines
  • Parental cell line: RPE1
  • Organism: Human
  • Tissue: Eye
  • Model: Reporter
  • Conditional: No
  • Description: Somatic knock-in reporter cell line with endogenous expression of Mad2 fused to the yellow fluorescent protein Venus (Venus-Mad2 fusion protein). Different cyclins exhibit distinct expression and degradation patterns which contribute to the temporal coordination of each mitotic event. This cell line functions as a cell cycle reporter across the metaphase-to-anaphase transition of the cell cycle. See also the Cyclin A2 and Cyclin B1 versions of these cell lines for a broader analysis of the different phases of the cell cycle. This cell line is an endogenous reporter for Mad2 levels and activity.
  • Research area: Cell biology
  • Production details: To generate the cell line recombinant adenovirus-associated virus (rAVV)-mediated gene targeting was used to fuse the yellow fluorescent protein (Venus) ORF to the 5' of exon 1 of one allele of the MAD2L1 gene. The Mad2 was tagged at its N-terminus because a GFP-Mad2 fusion protein had previously been demonstrated to be functional in Drosophila.

  • For Research Use Only

Target Details

  • Target: Mad2

Application Details

Handling

  • Format: Frozen
  • Growth medium: F12/DMEM (Sigma: D6241) + 2mM GlutaMAX?(Invitrogen), 10% Foetal Bovine Serum (FBS) + 0.348% sodium bicarbonate; 37?‚°C; 5% CO2.
  • Storage conditions: Liquid Nitrogen
  • Shipping conditions: Dry ice
  • Mycoplasma free: Yes
  • Biosafety level: 1

Documentation

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References

  • •  Collin et al., 2013. Nat Cell Biol. 15(11):1378-85. PMID: 24096242