This protocol can be used to generate stable cell lines expressing a gene of interest from an integrated lentiviral vector.
Cell line generation protocol.
Infection of human b cells with ebv in vitro results in their immortalization and the resulting cell lines are named lymphoblastoid cell lines lcls in these cells ebv establishes mainly a latent infection characterized by the expression of a limited number of viral proteins.
Identify single clones by limited dilution.
Stable cell line generation protocol.
This protocol can be used to generate stable cell lines expressing a gene of interest from an integrated lentiviral vector.
Atcc protocol was followed for subculturing.
This is usually the first step before moving to stable cell line development in order to be able to ensure that all antibody features meet the requirements in cho based productions.
Culture conditions for generation of stable cell lines as for transient transfection experiments culture conditions passage number split rhythm etc of your selected cell type are very important for the generation of stably transfected cell lines.
Unlike the short term protein expression observed using transient transfection methods generating cell lines using lentiviral vectors enables long term protein expression studies.
Maintenance of cell line before spheroid generation.
For optimal results we recommend following the cell culture recommendations of the supplier.
For stable cell line generation services vectorbuilder defines estimated turnaround as the time from production initiation to completion.
Our xtencho tm cell line together with our in house xten protocol is your best chance at overcoming your difficult to express protein challenge.
Production completion is achieved once the final cell line passes qc and is ready to be shipped out.
After thawing from liquid nitrogen sw480 cells were maintained in nunclon delta t25 cell culture flasks in gibco dmem medium supplemented with 10 gibco fbs and 1 pen strep for 2 passages before seeding for spheroid generation.
The protocol for generating stable cell lines requires several steps as shown below.
This protocol is specific for the generation of a monoclonal cell line that resistance to antibiotics g418 neomycin.
The end result that you are looking for is a population of cells in which 100 of cells are expressing your fusion protein.
Unlike the short term protein expression observed using transient transfection approaches generating cell lines using lentiviral vectors enables long term protein expression studies.
Transfect cells with desired plasmid construct s select and expand stable polyclonal colonies.
Turnaround time does not include waiting time for any customer supplied materials e g.
