c*****o
发帖数: 18 | 1 Hi folks,
I try to purify a peptide(AMP) from the bacteria broth culture. It looks
like that so much sugar from the broth tightly bound with the protein even
after HPLC and centricon pufication. I still find the dark sugar color in
the active fraction. and this is a big problem for the later SDS-PAGE
running since suger blocks the gel pore, I can not get a band......always
smear. Does somebody have experience to romove suger?
thanks for your time and help. |
s*******e
发帖数: 1010 | |
K******S
发帖数: 10109 | 3 Are you sure those are sugar?
【在 c*****o 的大作中提到】
: Hi folks,
: I try to purify a peptide(AMP) from the bacteria broth culture. It looks
: like that so much sugar from the broth tightly bound with the protein even
: after HPLC and centricon pufication. I still find the dark sugar color in
: the active fraction. and this is a big problem for the later SDS-PAGE
: running since suger blocks the gel pore, I can not get a band......always
: smear. Does somebody have experience to romove suger?
: thanks for your time and help.
|
c*****o
发帖数: 18 | 4 it should be the sugar. The culture broth contains high ammount of glucose(
20g/L), so after autoclave the broth color is dark brown. active fraction
eluted from HPLC has even more dark color.
it also looks like difficult to diaysis.... |
s******9
发帖数: 283 | 5 "so much sugar from the broth tightly bound with the protein even
after HPLC and centricon pufication"
Sounds wired... How did you do your HPLC? |
M*******a
发帖数: 15 | 6 no idea what happened. could it be possible to use Ca2+ for competing
binding and replace glucose?
【在 c*****o 的大作中提到】
: Hi folks,
: I try to purify a peptide(AMP) from the bacteria broth culture. It looks
: like that so much sugar from the broth tightly bound with the protein even
: after HPLC and centricon pufication. I still find the dark sugar color in
: the active fraction. and this is a big problem for the later SDS-PAGE
: running since suger blocks the gel pore, I can not get a band......always
: smear. Does somebody have experience to romove suger?
: thanks for your time and help.
|
n***w
发帖数: 2405 | 7 why you add glucose to your culture medium? to lower the background? |
