Stable Cell Line

Adherent mammalian cells are sensitive to concentrations of 2 5 μg ml while cells in suspension are sensitive to concentrations as low as 0 5 2 μg ml.

Stable cell line.

Transfect cells with expression vector. Focused on our comprehensive expertise in the creation of stable cell lines. Selection markers can be delivered using the same plasmid that contains the gene of interest in cis or on a separate plasmid in trans that needs to be co transfected with the plasmid containing the gene of interest. Unlike the short term protein expression observed using transient transfection approaches generating cell lines using lentiviral vectors enables long term protein expression studies.

Stable cell line generation is made possible by the use of positive selection markers such as g418 hygromycin b puromycin resistance etc. Selection markers can be delivered using the same plasmid that contains the gene of interest in cis or on a separate plasmid in trans that needs to be co transfected with. The following day replace the standard media with media containing g418. This protocol can be used to generate stable cell lines expressing a gene of interest from an integrated lentiviral vector.

To set up for the stable cell line selection split cells and transfect with your construct. The target cell line was a readily available inducible cell line. The amount of g418 required to kill cells. We are happy to customize services as needed and discuss your experimental details to ensure an accurate quote.

There are two types of stable cell lines. Stable expression of target gene in cell lines overcomes the low transfection efficiency of transient expression and produces more target proteins. By building your your project in this way you can be guaranteed of support throughout the development process giving you unique flexibility. Stable cell line generation is made possible by the use of positive selection markers such as hygromycin g418 geneticin zeocin and blasticidin antibiotic resistance.

Stable cell line services is a division of altogen labs inc. Antibodies were identified and validated for this purpose. Puromycin has a fast mode action causing rapid cell death even at low antibiotic concentrations allowing the generation of puromycin resistant stable cell lines in less than one week. The screening procedure was determined to be western blot analysis of the inducible expressed protein.

Our stable cell line development service is composed of several milestones go no go steps that are accompanied by a detailed report and discussed with our expert team.