Overexpression of survivin in these stable transfectants
was verified by immunoblotting.
were transferred and maintained in DMEM supplemented with 10% FBS and 400 mg/ml G418.
We transfected Chinese hamster ovary (CHO) cells (American Type Culture Collection) with the human EL-c-myc/pHBAP-3-neo plasmid (10) and selected stable transfectants
with 500 [micro]g/mL G418 (Invitrogen).
The stable transfectants
were established and cultured in antibiotic-free DMEM for at least two passages before performing experiments.
do not express sufficient levels of fluorescence for spectrofluorometric measurement on micro plate.
were still expressing the marker gene, more than 30 days post transfection, when the study was terminated.
The broad dynamic range of the Guava Express assay provides the precise discrimination necessary for studying cell receptor up/down-regulation, screening transfectants
, evaluating whole blood lymphocyte populations, and many other applications.
ELISA assay showed that the transfectants
secreted hTPO protein in media at the level of 774 pg/ml (pBCTPOKI-6) and 1,867 pg/ml (pBCTPOKI-10), respectively (Figure 1C).
After stable transfection of HEK-293 cells with the a, [beta], and [gamma] subunits of the human Fc[epsilonRI, we identified transfectants
expressing the functional receptor by use of FRCS using an [alpha] chain-specific antibody (Fig.
Mouse epidermal JB6 Cl41 cells and their stable transfectants
were cultured in Eagle's minimal essential medium (MEM; Calbiochem, San Diego, CA) supplemented with 5% fetal bovine serum (FBS), 1% penicillin/streptomycin, and 2 mM L-glutamine (Life Technologies, Inc.
In addition to the parental cell line preparation, which produced a moderate amount of BCAR1, the study included the transfectant
4A12, which overproduced BCAR1 (1), and transfectants
that overproduced FLAG-tagged (Stratagene Europe) BCAR1 (FBCAR1), HEF1, and a FLAG-tagged hybrid molecule consisting of the first 63 amino acid residues of BCAR1 and residues 64-834 of HEF1 (F-Hybrid).
This assay uses MCF7-AR1 cells, which are stable transfectants
of MCF7 cells expressing the wild-type human androgen receptor (Szelei et al.