Description
GenJet™ DNA In Vitro
Transfection Reagent for MB231 is pre-optimized for transfecting MDA-MB231 cells.
The MDA-MB-231 breast cancer cell line was obtained from a pleural
effusion of a 51-year-old white female in October 17, 1973, by Dr. R. Calleau at
M. D. Anderson Cancer Center. The patient underwent a mastectomy and
radiotherapy prior to the removal of the breast cancer specimen. After 52
passages, the karyotype displayed chromosome numbers ranging between 52 and 62,
where the modal chromosome number was 57. With epithelial-like morphology,
the MDA-MB-231 breast cancer cells appear phenotypically as spindle shaped
cells. In vitro, the MDA-MB-231 cell line has an invasive phenotype. It
has abundant activity in both the Boyden chamber chemoinvasion and chemotaxis
assay. The MDA-MB-231 cell line is also able to grow on agarose, an indicator of
transformation and tumorigenicity, and displays a relatively high colony forming
efficiency.
The "Shaved Cell
Transfection" protocol was optimized to achieve maximal transfection efficiency. Refer to the following optimal transfection conditions for maximal transfection
efficiency on MB231 cells. GenJet™ reagent, 1.0 ml, is sufficient for 300 transfections in 24 well plates or 150 transfections in 6 well plates.
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Summary of
Optimal Shaved CellTransfection Conditions:
Cell culture
medium
Confluency
of cell on the
day of shaving
Cell density for plating
GenJet™ (µl) : DNA (µg) Ratio
Diluent for DNA and Transfection Reagent
Incubation Time to Form GenJet™/DNA
Complex
Incubation time of shaved cells with transfection complex
Maximal Efficiency
Transfection
Results:
Reporter Gene
Plasmid
Efficiency (GFP %) |
DMEM with 4.5 g/L glucose, 10% FBS
95~100%
1.3x10^5 cells per square centimeter
4:1
Serum-free DMEM with 4.5 g/L glucose
12 minutes at RT
20 minutes at 37 degree
48 hours
EGFP
pEGFP-N3 (CMV promoter)
70% |
Storage Condition
Store at 4 °C. If stored properly, the product is stable for 12 months or longer
A Picture Showing Transfection Efficiency of GenJet™ Reagent on MDA-MB231 Cells.
MDA-MB231 cells
were grown per ATCC recommended culture medium and transfected with pEGFP-N3 by
GenJet™. The efficiency was checked 24 hours post transfection
Data Sheet
