HT-29/NF-kB Reporter (Luc) stable cell line荧光稳转报告细胞株 BioVector NTCC质粒载体菌种细胞基因保藏中心

HT-29/NF-kB Reporter (Luc) stable cell line荧光稳转报告细胞株

Product description
HT-29/NF-kB Reporter (Luc) cells are derived from the human colon cancer cell line HT-29 by
stably integration of a NF-kB firefly luciferase reporter construct. HT-29 cells have been used in
cancer research and drug development. HT-29/NF-kB Reporter (Luc) cells express firefly
luciferase under the control of the NF-kB response elements, can be used for in vitro assays
and in vivo imaging.

Figure 1. HT-29/NF-kB Reporter (Luc) cells in response to hTNFα.
The luminescence intensity of ~5000 cells was detected by Bright-GloTM luciferase Assay System
(Promega, Cat E2610).
Cell line description
Organism: Homo sapiens (human)
0.00E+00
1.00E+04
2.00E+04
3.00E+04
4.00E+04
5.00E+04
6.00E+04
7.00E+04
8.00E+04
9.00E+04
1.00E+05
untreated 20 ng/ml hTNFα
Relative luminometer units
Tissue: Colon
Morphology: epithelial
Disease: Adenocarcinoma; Colorectal
Culture Properties: Adherent
Biosafety Level: 2
Medium
1. Complete culture medium: DMEM, 10% fetal bovine serum (FBS)
2 µg/mL of puromycin may be added to the culture medium. Puromycin should not be
added until a culture has been well established from the thawed cells.
2. Freeze medium: FBS with 6% DMSO
Culture procedure
Thawing of frozen cells
1. Thaw the frozen cryovial by gentle agitation in a 37 °C water bath in 1-2 minutes.
2. Remove the cryovial from the water bath as soon as the contents are thawed, and
decontaminate by wiping with 70% ethanol.
3. Transfer the thawed cell suspension to a centrifuge tube containing 10 ml of Complete
culture medium, centrifuge at 500 g for 5 minutes.
4. Remove the medium by aspiration, resuspend the cells with 10 ml of the Complete
culture medium by gently pipetting up and down.
5. Transfer the cells to a 10 cm cell culture dish.
6. Place the cells in a 37°C incubator with 5% CO2.
Sub-culturing
Volumes are given for a 10 cm cell culture dish. Increase or decrease the amount of
dissociation medium needed proportionally.
1. Remove the medium by aspiration.
2. Briefly rinse the cell layer with 1xDPBS to remove all traces of serum that
contains trypsin inhibitor.
3. Add 1 ml of Trypsin-EDTA (0.05%) solution to the dish and observe cells under
an inverted microscope until cell layer is dispersed.
4. Add 4 ml of complete growth medium and aspirate cells by gently pipetting.
5. Add appropriate aliquots of the cell suspension to new culture vessels. Incubate
cultures at 37°C with 5% CO2.

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