stable cell line questions( cell type matter) - (Nov/10/2006 )
I am recently planning to build a stable cell line which will overexpressing the protein I am interested.
1. My first question is whether I could use whatever types of immortalized cell lines availabe for this?
2. I have actually detected the induction of endogenerous protein ( protein XX) in Mes 23.5 cells ( a neuronal cell line), can I use this cell line for builiding stable cell line which will overexpressing protein XX? Does the cell line I choose have to be protein XX-deficient at the first place?
I am a starter for stable transfection. Any comments or suggestions will be highly appreciated 
1. My first question is whether I could use whatever types of immortalized cell lines availabe for this?
2. I have actually detected the induction of endogenerous protein ( protein XX) in Mes 23.5 cells ( a neuronal cell line), can I use this cell line for builiding stable cell line which will overexpressing protein XX? Does the cell line I choose have to be protein XX-deficient at the first place?
I am a starter for stable transfection. Any comments or suggestions will be highly appreciated
what is the aim of overexpression? so the type of cell may be interesting; your chosen cell type must not be xx-deficient beside you like to substitute one isoform against another to show the effects of different isoforms;
establishing a stable cell line may not succeed in any case; f.i. some potent regulators, kinases or receptors may be suppressed as their overexpression is cytotoxic
hi
protein overexpression for purification matter should be done in classical HeLa or 293cells in some cases as they're really easy to grow and to break for purification. You should use btw a strong promoter like CMV one.
After that point, if your aim is to overexpress a part of your protein, and study if it's ok in living cells that are knocked down for the whole protein, yes you should choose the Mes 23.5.