c**********n
发帖数: 177 | 1 小弟最近在做SPR，用的是biacore T100，sensorgram上得到的结果一直很诡异。在
assocition和dissociation的地方都有很高的信号（请看附图）。请大神们帮忙分析下
原因，因为是第一次做SPR经验不足，看manual也看不出个所以然。
做的是两个蛋白的结合，其中一个蛋白有biotin tag，被铆钉在SA chip上，另一个蛋
白过flow cell，flow cell 1没有铆钉任何蛋白作为blank，附图的结果是flow cell 4
subtracts flow cell 1。如果大神还要什么信息，我会尽量补充，先谢过各位。 |
d******4
发帖数: 1 | 2 buffer 的 composition或者PH和蛋白里面的不一样。 |
c**********n
发帖数: 177 | 3 多谢啊，我的analyte的buffer用的是50mM Tris PH 8.0, running buffer 用的是PBS
PH 7.4+0.5% P20，我做个buffer exchange试试看。
【在 d******4 的大作中提到】
: buffer 的 composition或者PH和蛋白里面的不一样。
|
s********n
发帖数: 248 | 4 二楼说的对。buffer 的PH值或者盐浓度不一样。
这问题我也遇到过。 |
c**********r
发帖数: 138 | 5 I disagree. If it's the discrepancy between running buffer and sample buffer
, there ought to be a lasting phase of high signal. Instead, we only see
spikes here. It is Injection spikes. Usually they are caused by mechanical
disturbance of injection. You can easily get rid of them using scrubber. |
c**********n
发帖数: 177 | 6 You mean disrobe before the run? Thanks for your reply!
buffer
【在 c**********r 的大作中提到】
: I disagree. If it's the discrepancy between running buffer and sample buffer
: , there ought to be a lasting phase of high signal. Instead, we only see
: spikes here. It is Injection spikes. Usually they are caused by mechanical
: disturbance of injection. You can easily get rid of them using scrubber.
|
