Get rib of albumin - (Jan/24/2008 )
I am going to purify a His-tagged protein from cell culture medium. But I ran a SDS-PAGE and found that albumin existed as a thick band in the gel. Will the albumin affect my purification process. The column I will use is from GE healthcare (His-trap FF), will albumin bind to the column?
-Durandal-
QUOTE (Durandal @ Jan 25 2008, 11:44 AM)
I am going to purify a His-tagged protein from cell culture medium. But I ran a SDS-PAGE and found that albumin existed as a thick band in the gel. Will the albumin affect my purification process. The column I will use is from GE healthcare (His-trap FF), will albumin bind to the column?
Google "albumin removal" and you'll find a pile of companies make resins for removal of albumin (typically for 2D gels with low-level proteins).
-swanny-
albumin should have no effect on the histrap column.
-mdfenko-
QUOTE (swanny @ Jan 25 2008, 05:55 AM)
QUOTE (Durandal @ Jan 25 2008, 11:44 AM)
I am going to purify a His-tagged protein from cell culture medium. But I ran a SDS-PAGE and found that albumin existed as a thick band in the gel. Will the albumin affect my purification process. The column I will use is from GE healthcare (His-trap FF), will albumin bind to the column?
Google "albumin removal" and you'll find a pile of companies make resins for removal of albumin (typically for 2D gels with low-level proteins).
we use Beckman Coulter colummns for albumin depletion for 2D gels, they work fine althought I think they're a bit expensive
-Pumuki-
It is a smart idea to adapt your cells in serum-free condition. It takes about 3-15 days. You will feel much comfortable with your purification. Let me know if there is any technical difficulty.
QUOTE (Pumuki @ Jan 31 2008, 10:58 AM)
QUOTE (swanny @ Jan 25 2008, 05:55 AM)
QUOTE (Durandal @ Jan 25 2008, 11:44 AM)
I am going to purify a His-tagged protein from cell culture medium. But I ran a SDS-PAGE and found that albumin existed as a thick band in the gel. Will the albumin affect my purification process. The column I will use is from GE healthcare (His-trap FF), will albumin bind to the column?
Google "albumin removal" and you'll find a pile of companies make resins for removal of albumin (typically for 2D gels with low-level proteins).
we use Beckman Coulter colummns for albumin depletion for 2D gels, they work fine althought I think they're a bit expensive
-HCscientist-