secretory proteins - recombinant proteins (Jan/08/2009 )
Hi,
My lab is running on a loww budget. As much as I love to order a recombinat protein, I've been told o purify it inlab. I've cloned my proten in a mammalian expression vector (pcDNA3.1) and its got a his tag aand a myc tag. Protein size= 28kDa.
1) The protein of interest is secretory. Thus, do you think I shall get a better purity if I work with the supernatant of a stable cell line overexpressing my protein?
2) I have to clone, stable transfect and purify 3 proteins and thus all efficient methods are appreciated!!
-Sarwat-
QUOTE (Sarwat @ Jan 8 2009, 10:11 AM)
Hi,
My lab is running on a loww budget. As much as I love to order a recombinat protein, I've been told o purify it inlab. I've cloned my proten in a mammalian expression vector (pcDNA3.1) and its got a his tag aand a myc tag. Protein size= 28kDa.
1) The protein of interest is secretory. Thus, do you think I shall get a better purity if I work with the supernatant of a stable cell line overexpressing my protein?
2) I have to clone, stable transfect and purify 3 proteins and thus all efficient methods are appreciated!!
My lab is running on a loww budget. As much as I love to order a recombinat protein, I've been told o purify it inlab. I've cloned my proten in a mammalian expression vector (pcDNA3.1) and its got a his tag aand a myc tag. Protein size= 28kDa.
1) The protein of interest is secretory. Thus, do you think I shall get a better purity if I work with the supernatant of a stable cell line overexpressing my protein?
2) I have to clone, stable transfect and purify 3 proteins and thus all efficient methods are appreciated!!
Well better purity
you wil stil have to purify it onto a Nickel column or something similar anyway because you will get all the proteins from the media (FCS, Albumin etc..) If you talk about integrity of teh protein hard to predict anyway 
-Jipes-