r****1
发帖数: 2299 | 1 做了一个PEPTIDE，里面有两个CYS，想让这两个CYS FORM DISULFIDE BRIDGE。应该如
何做才能加速这个反应，而且还要避免INTERMOLECULAR CROSS LINK而形成DIMER。 |
h*****e
发帖数: 1330 | 2 看你挺眼熟，浓度稀释不知道能不能work，那个老印不是搞这个的么
【在 r****1 的大作中提到】
: 做了一个PEPTIDE，里面有两个CYS，想让这两个CYS FORM DISULFIDE BRIDGE。应该如
: 何做才能加速这个反应，而且还要避免INTERMOLECULAR CROSS LINK而形成DIMER。
|
B******1
发帖数: 9094 | 3 Theoretically, you need to know the reaction rate for intramolecular and
intermolecular disulfide-formation. Then you can decide how dilute you want
to go to maximize the yield of intramolecular disulfide bond formation.
Running the same reaction under different concentrations and temperatures,
among other things, would give you some hard data to work with. Hopefully
after a few rounds you would be able to find out the optimal conditions.
If your goal is simply to have a few mg of the desired |
k****o
发帖数: 728 | 4 Considering peptide is too small, is SDS-PAGE capable of separating 3000 Da
peptide from 6000 Da peptide ?
want
to
【在 B******1 的大作中提到】
: Theoretically, you need to know the reaction rate for intramolecular and
: intermolecular disulfide-formation. Then you can decide how dilute you want
: to go to maximize the yield of intramolecular disulfide bond formation.
: Running the same reaction under different concentrations and temperatures,
: among other things, would give you some hard data to work with. Hopefully
: after a few rounds you would be able to find out the optimal conditions.
: If your goal is simply to have a few mg of the desired
|
r****1
发帖数: 2299 | 5 ok, i see. HPLC probably can separate the right compound. and for the
reaction, just leave the peptide in solution for couple of days is enough? |
h*****e
发帖数: 1330 | 6 如果产物稳定，应该可以有个什么方法detect whether the reaction is done or not
. -SH和-S-S-还是有差别 |
h*****e
发帖数: 1330 | 7 还有自己找找文献，NYU查文献那么方便不好好利用挺可惜 |
l****y
发帖数: 398 | 8 DTNB
【在 r****1 的大作中提到】
: 做了一个PEPTIDE，里面有两个CYS，想让这两个CYS FORM DISULFIDE BRIDGE。应该如
: 何做才能加速这个反应，而且还要避免INTERMOLECULAR CROSS LINK而形成DIMER。
|
t*******e
发帖数: 440 | 9 DTNB and NBD assay
not
【在 h*****e 的大作中提到】
: 如果产物稳定，应该可以有个什么方法detect whether the reaction is done or not
: . -SH和-S-S-还是有差别
|
t*******e
发帖数: 440 | 10 DTNB and NBD assay
not
【在 h*****e 的大作中提到】
: 如果产物稳定，应该可以有个什么方法detect whether the reaction is done or not
: . -SH和-S-S-还是有差别
|
U*****e
发帖数: 40 | 11 0.2 mg per ml in 100 mM Phosphate buffer pH 7.4, air oxidation over night at
room temperature, check the change of HPLC retention time and mass |
b*****r
发帖数: 203 | 12 pH 值影响应该也挺大吧！
【在 r****1 的大作中提到】
: 做了一个PEPTIDE，里面有两个CYS，想让这两个CYS FORM DISULFIDE BRIDGE。应该如
: 何做才能加速这个反应，而且还要避免INTERMOLECULAR CROSS LINK而形成DIMER。
|
P***e
发帖数: 804 | 13 Pt catalyst:
http://www.ncbi.nlm.nih.gov/pubmed/12127546
Convenient synthesis of human calcitonin and its methionine sulfoxide
derivative
【在 r****1 的大作中提到】
: 做了一个PEPTIDE，里面有两个CYS，想让这两个CYS FORM DISULFIDE BRIDGE。应该如
: 何做才能加速这个反应，而且还要避免INTERMOLECULAR CROSS LINK而形成DIMER。
|
P***e
发帖数: 804 | 14 Run LC-MS to monitor disulfide peptide change over time.
If no MS connected to LC, collect fractions then run MALDI.
In terms of purification, HPLC is the way to go.
not
【在 h*****e 的大作中提到】
: 如果产物稳定，应该可以有个什么方法detect whether the reaction is done or not
: . -SH和-S-S-还是有差别
|
