Dear all,
sorry for the slightly off topic post,
I have 2 proteins that have been shown to interact, by multiple groups, and
by multiple techniques - namely ELISA, SPR and DPI.
The Kd of the interaction as determined by SPR is on the order of 1 nM.
I would very much like to crystallise this
Dear Dave,
Indeed, an interaction in the nM range is strong, but its dynamic is
important. If the SPR sensorgram, if published, looks like a 'crenel',
this indicates high association/dissociation rates. In that case, your
complex can dissociate during the GF run.
You can try crystallisation
Hey David,
the 1 nM Kd by SPR sounds fishy to me - did you do these measurements yourself ?
Unless this is an antibody-protein complex or nanobody-protein complex or
protein-small molecule complex, the value is too low (for a normal PPI, I would
expect 50 -500 nM). Possible explanation for the
Subject: [ccp4bb] Isolation of protein-protein complexes.
Dear all,
sorry for the slightly off topic post,
I have 2 proteins that have been shown to interact, by multiple groups, and by
multiple techniques - namely ELISA, SPR and DPI.
The Kd of the interaction as determined by SPR is on the order
Hi all,
Thanks for the incredibly fast and helpful responses, both on and off list.
To provide some further information. I have tried the SEC with several
buffers, including buffers identical to the SPR and DPI experiments.
To-date all SEC and SPR experiments have been conducted at room
@JISCMAIL.AC.UK
*Subject:* [ccp4bb] Isolation of protein-protein complexes.
Dear all,
sorry for the slightly off topic post,
I have 2 proteins that have been shown to interact, by multiple groups,
and by multiple techniques - namely ELISA, SPR and DPI.
The Kd of the interaction as determined
that was in the
buffer
used for the other binding studies, but not in your SEC buffer?
JPK
*From:* CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] *On Behalf
Of *David
Briggs
*Sent:* Tuesday, January 21, 2014 10:52 AM
*To:* CCP4BB@JISCMAIL.AC.UK
*Subject:* [ccp4bb] Isolation of protein