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Stable Cell Lines

Watsonbio’s stable cell line service can build your cell lines based on your needs, either for protein expression or assay development. You can choose the best way to generate your stable cell line.

Stable transfection and screening

Development of a stable cell line based on CHO for production of recombinant proteins, or any cell lines provided by clients. Wastonbio provides CHO cell line based on proprietary expression vector, high throughput cell screening technologies.

Stable transfection and screening

  • Gene synthesis and plasmid generation
  • Transient expression evaluation
  • Transfection and clone screen

Lentivirus stable cell line (overexpress or shRNA knockdown)

  • Construct your gene into our lentiviral vector with desired selection marker
  • Generate lentivirus
  • Transduce the cell line of your interest
  • Select the stably transduced cells

293T/TetR-Flt3 cell lines were cultured in the presence or absence of 3 ug/mL tetracycline for 72 hrs. After the induction, whole cell extracts (WCE) were isolated. Thirty microgram total proteins were loaded for SDS-PAGE isolation. Polyclonal antibody against Flt3 and GAPDH antibody were used for Western Blot detection.

Isogenic Cell Lines

Genetically engineered cells generated by randomly integrating DNA into a cell’s genome have the problems encountered ranging from unpredictable expression levels to safety concerns. Using recombinase-mediated chromosome engineering (FLP/FRT recombination system), single-copy integration of foreign DNA fragments can be achieved at predetermined chromosomal loci in the genome to generate isogenic cell lines. We provide customized services based on this technology to various human cell lines with a rapid turnaround time about 8-10 weeks

Stable Expression of GPBAR1 in 293T/FLP cells. Fifty microgram whole cell extracts were used for the Western Bolt analysis. GAPDH was served as a loading control.

293T-FLP cells with stable expression of GPBAR1 receptor were incubated with Forskolin and indicated amounts of agonist in induction buffer. cAMP levels alterations were measured using a luminescence-based assay. Each point represents three data points, the error bars represent SEM.

Deliverables

  • Two vials of stable cells and final report
  • Validation the genomic integration of gene via genomic PCR
  • Validation the high-expression clone by Western Blot (if applicable)
  • Any specific assay based on request