Enzyme-Free Cell Dissociation Solution

NIH 3T3 cells transfected with an expression vector (pMV7) containing a murine leukemia virus long terminal repeat which serves as the promoter for expression of the rat serotonin 5HT1c cDNA and an independent expression cassette encoding neomycin phosphotransferase. Cells were dissociated with ENZYME FREE CELL DISSOCIATION solution and loaded with the Ca++ sensitive dye Indo-1. Activation of 5HT1c receptors elevates intracellular Ca++ concentration. Changes in the level of intracellular free Ca++ were monitored with a flow cytometer. The ligand binding and second messenger activation functions are intact and demonstrate 5HT1c specific responses.

HELA cells transfected with an expression vector (pMV7, described above) containing the cDNA encoding human CD4. Cells were dissociated with ENZYME-FREE CELL DISSOCIATION SOLUTION and incubated with a monoclonal antibody for CD4 or CD8. The cells were then washed and incubated with a FITC conjugated secondary antibody. The CD4 proteins expressed on the cell surface were specifically stained by anti-CD4 and did not stain with any anti-CD8 or secondary antibody alone. These experiments demonstrate the antigenic and structural integrity of the CD4 proteins expressed on the surface of transfected HELA cells after dissociation with the ENZYME FREE CELL DISSOCIATION SOLUTION.