Dear Patrick,
I did use microseeding but for a different ligand. Will try with this as
well.
Thank you
REgards
Kavya
On 2023-02-07 16:20, Patrick Shaw Stewart wrote:
>> As to why previously in a very similar condition you did get your desired
>> protein plus (other) ligand
>
> Kavya,
>
> As to why previously in a very similar condition you did get your desired
> protein plus (other) ligand
Kavya, did you use microseeding? That's the way to get consistent results.
Since you're changing the ligand I suggest you go back and run a few random
screens (with crushed crystals of
Dear Ben,
It was a single crystal that was mounted. I did use IZIT dye, did not
trun blue much. Probably its a TCEp crystal. But I did try a negative
control without protein+ligand and rest everything same, the drop was
clear!
Thank you
Regards
Kavya
On 2023-02-06 22:49, Ben Bax wrote:
SENT: Saturday, February 4, 2023 14:32
> TO: CCP4BB@JISCMAIL.AC.UK
> SUBJECT: Re: [ccp4bb] [EXT] Re: [ccp4bb] Regarding the diffraction image
>
> PS it's probably not a salt crystal...TCEP is not a salt, your ligand I
> presume is also not a salt, a small cleaved peptide ne
assisted by anomalous P
(or Zn?) signal...
Have fun, BR
From: CCP4 bulletin board On Behalf Of Mark J. van Raaij
Sent: Saturday, February 4, 2023 14:32
To: CCP4BB@JISCMAIL.AC.UK
Subject: Re: [ccp4bb] [EXT] Re: [ccp4bb] Regarding the diffraction image
PS it’s probably not a salt crystal…TCEP
Dear Mark,
Thanks for the clarification.
Regards
Kavya
On 2023-02-05 04:02, Mark J. van Raaij wrote:
> PS it's probably not a salt crystal...TCEP is not a salt, your ligand I
> presume is also not a salt, a small cleaved peptide neither. As to why
> previously in a very similar
PS it’s probably not a salt crystal…TCEP is not a salt, your ligand I presume
is also not a salt, a small cleaved peptide neither. As to why previously in a
very similar condition you did get your desired protein plus (other) ligand
crystal, it just means the molecule (TCEP') crystallises in a
the unit cell in the c direction is quite long, 49 Å, this gives the relatively
close spots in one direction.
Mark J van Raaij
Dpto de Estructura de Macromoleculas, lab 20B
Centro Nacional de Biotecnologia - CSIC
calle Darwin 3
E-28049 Madrid, Spain
tel. +34 91 585 4616 (internal 432092)
Section
Dear all,
Sorry for the confusion created, I did not mean that a protein would
have fit in the small unit cell. My question was -
1. Why are there closely spaced spots arising in salt crystal?
2. If TCEP could crystallize in the condition, I have got a protein
(same as this)+ligand
Dear Mark,
I did think it was salt, so I checked the same batch of protein which
went for crystallization by running a gel, it was intact, no cleavage.
My doubts arose because of two things
1. I crystallized the same protein with another ligand, in very similar
condition (10% PEG3350, 50mM Zn
Hi Jessica,
I am quite sure the protein cannot be fit in this unitcell. I was just
curious why the diffraction has closely spaced spots.
Thanks
On 2023-02-04 00:02, Jessica Bruhn wrote:
> Hi Kavya,
>
> As others have mentioned, the unit cell is too small to contain your protein.
> With
Dear Thomas,
Interestingly, I had crystallized the same protein with another ligand
before with the same condition except that it had sodium citrate in
addition. I was able to collect the data for this and it was a
protein-ligand complex, which could be seen in the density. So I was
speculating
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