TCF/LEF Luciferase Reporter Stable Cell line (For Research Use Only)

 

Catalog Nunber

#STC003

Source

Human dermal papilla cell line (catalog number #STC001)

Introduction

The Wnt signaling pathway is important to both embryonic development and tumorigenesis. b-catenin, the central component of the pathway, functions as a co-factor of the TCF/LEF family of transcription factors. Together, they activate transcription of Wnt target genes by binding to the promoters of downstream target genes involved in cell proliferation, survival, and migration. STEMORE has established a TCL/LEF luciferase reporter stable cell line that has been stable transfected with pGreenFire-TCF/LEF reporter vector, which contains 7 repreats of TCF/LEF binding sites, a minimal promoter upstream of the firefly luciferase coding region. Therefore, the cell line can be used as a reporter system for monitoring the activation of b-catenin triggered by stimuli treatment, enforced gene expression and gene knockdown.

Product description

STEMORE has developed TCL/LEF luciferase reporter stable cell line by infecting TCF/LEF reporter vector. The puromycin resistant clones were subsequently screened for LiCl and Wnt3a-induced luciferase activity. The cell line can be used as a reporter system for monitoring the activation of b-catenin triggered by stimuli treatment, such as Wnt3a, LiCl, and gene overexpression and gene knockdown.

Materials provide

One vial of 1 ´ 106 cells. Store the frozen cells in liquid nitrogen until you are ready thaw and propagate them.

Handling and Storage

The cells should be cultured in complete DMEM medium [In 450 mL of high-glucosed DMEM, add 50 mL FBS (10% final) and 5 mL Penicillin/Streptomycin (1% final)].

Initial Culture Procedure

  1. Quickly thaw cells in a 37°C water bath. Remove from the as soon as the vial is thawed.
  2. Transfer cells to 100 mm2 dish containing 10 ml of complete growth media.
  3. Gently rock the dish to ensure the cells are mixed well in the media.
  4. Place the dish with cells in a humidified incubator at 37°C with 5% CO2.
  5. After cells adhere (wait at least 4 hours to overnight), replace media with fresh complete growth medium.

Subculture Procedure

  1. Subculture cells when density reaches 90~100% confluency.
  2. Carefully remove the culture media from cells by aspiration.
  3. Rinse cells with PBS, being careful to not dislodge attached cells. Then remove PBS by aspiration.
  4. Add 1~2 mL trypsin-EDTA solution.
  5. Incubation with trypsin for 2~5 minutes (or until detached). Confirm detachment by observation under the microscope.
  6. Add 5~10 mL of pre-warmed complete growth media and gently pipet up and down to break the clump.
  7. Passage cells in 1:3 to 1:5 ratio when they reach 90% confluency.