Description

GenJet™ DNA In Vitro Tranfection 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.
T
he "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.

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 c
entimeter
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 CellsMDA-MB231 cells were grown per ATCC recommended culture medium and transfected with pEGFP-N3 by GenJet™. The efficiency was checked 24 hours post transfection