NCI-H292 Luciferase Reporter Cell Line

Frequently Asked Questions

• What is the recommended culture medium for the NCI-H292-Luc cell line?

  The recommended culture medium is 90% RPMI-1640 with 10% Fetal Bovine Serum (FBS). The use of penicillin/streptomycin (P/S) is optional, and it can be added after the cells have recovered and grown well after thawing.

• What is the procedure for thawing cryopreserved NCI-H292-Luc cells?

  Warm the complete culture medium in a 37°C water bath for 30 minutes.
  Transfer the cryovial from liquid nitrogen to -80°C and allow any residual nitrogen to volatilize.
  Thaw the vial in a 37°C water bath for about a minute while swirling gently.
  Once thawed, transfer the cells to a centrifuge tube with 6-7 mL of medium and centrifuge at 1100 rpm for 4 minutes.
  Resuspend the cells in fresh medium and transfer them to a T25 flask for incubation.

• How is luciferase activity validated in NCI-H292-Luc cells?

  The luciferase activity is validated using the Dual-Luciferase® Reporter Assay System. The intensity of the firefly luciferase reaction is measured using a Microplate Reader. The assay confirms strong luminescence, ensuring reliable performance of the luciferase gene expression system.

• What is the passage ratio for the NCI-H292-Luc cell line, and when should cells be passaged?

  Cells should be passaged when they reach 80%-90% confluence. The recommended passage ratio is between 1:2 and 1:4, depending on how quickly the cells reach confluence. For the first passage, a ratio of 1:3 is recommended.

• What precautions should be taken upon receiving cryopreserved NCI-H292-Luc cells?

  Upon receiving cryopreserved cells, they should be transferred immediately to liquid nitrogen or a -80°C freezer. Photos of the package, including dry ice and tubes, should be taken, and abnormalities like depleted dry ice or damaged cryovials should be reported within 24 hours.