Hi All,
I am purifying the biotinylated protein (cloned into the pET28a vector)
using Avidin beads. Since I need the protein for SPR but when I used the
purified protein to interact with Streptavidin coated onto the SPR chip.
There was no signal. Can anybody tell me why is it so or how can I make
Dear All
a heads up that the early bird registration, and closing date for student
bursaries, is Sunday 18th November 2018 i.e. this Sunday!
For more info see http://www.cvent.com/d/sgq8q6/1K?cpc=N7N8SK89V8N
Regards
Karen McIntyre
Scientific Computing Department - CCP4
Call Postdoctoral Fellowship Program at the Helmholtz Zentrum München, Munich,
Germany
Deadline November 30 , 2018
The PFP trains highly talented early-career scientists for leadership positions
in health and environment research.
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Hello,
you probably purified a contaminant. Do a blot with an anti-biotin
antibody or get electro-spray mass spectrometry done in order to
confirm the identity of your protein.
Wim
On 13/11/2018 11:13, Anamika Singh
wrote:
Hi All,
Hi Anamika,
- Have you double-checked that the sequence of your cDNA is correct and
includes the biotin acceptor peptide tag (BAP tag aka AviTag; GLNDIFEAQKIEWHE
in single-letter amino acid code)?
- Are you using a dedicated bacterial strain that over-expresses BirA enzyme?
This may not be
You don’t say so, but one assumes that you have a BAP tag on the protein, and
co-express a biotin ligase such as BirA?
Ed
T.A.Edwards Ph.D.
Associate Professor of Biochemistry
Deputy Head of School
_
Astbury Centre for Structural Molecular Biology
School of
Hi Anamika,
As far as I understood, the biotin in the elution
buffer is helping your protein to get stripped off from the Avidin column.
So, maybe you dialyze your purified protein (or run FPLC) and get rid of
biotin completely, before you load the protein on to strptavidin
Hi all,
I think I know why it is a division of 4 instead of 2 is involved in conversion
from VAX to IEEE now. Short answer: a 2 is in the exponent bits (bias of 128
instead of 127, visible), another 2 is hidden in the scientific notation.
I found this explanation+example on VAX F-float:
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There is an availability in the laboratory of Crina Nimigean for an
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It's also said here, at the end of file :
https://www.cs.auckland.ac.nz/~patrice/210LN/DR4.pdf
"add 1 to the left, with the binary point"
0.1.
From: CCP4 bulletin board on behalf of Zhijie Li
Sent: Tuesday, November 13, 2018 7:43 PM
To:
Hi Zhijie
It's definitely a factor 4. The code is in subroutine QTIEEE in the
Fortran source I mentioned previously at this line:
See line:
A(I)=((A(I)+SIGN(2,A(I)))/4.AND..NOT.MNAN).OR.MDN2
If you prefer it in C code it's in function vaxF2ieeeF in:
On Tuesday, November 13, 2018 1:06:01 PM PST Zhijie Li wrote:
> Hi Ethan,
> Thanks for the information. My guess is that in MTZ only F-float is expected,
> because it is the only 32bit form?
I do not remember exactly what was used for mtz files at that time.
It might have been REAL*4 or it
MTZ was always 32 bit floats for the main data, with ASCII headers at the end
Sent from my iPhone
> On 13 Nov 2018, at 21:29, Ethan A Merritt wrote:
>
>> On Tuesday, November 13, 2018 1:06:01 PM PST Zhijie Li wrote:
>> Hi Ethan,
>> Thanks for the information. My guess is that in MTZ only
On Tuesday, November 13, 2018 11:51:55 AM PST Zhijie Li wrote:
> If somebody is going to send these files by email, please send one to me too.
> Thanks in advance. I actually prefer to get a MTZ file because the miller
> indices would serve as good clues for understanding the encodings. Even
>> ah, nostalgia
Ah, "mantissa!" Haven't heard "mantissa" in decades...
Is there such a thing as a "praying mantissa?" Seems like there could be a good
geek joke about it.
JPK
> However all procedures I have seen use a division of 4, which is quite
> puzzling to me. A real data file
Hi Ethan,
Thanks for the information. My guess is that in MTZ only F-float is expected,
because it is the only 32bit form?
Zhijie
> On Nov 13, 2018, at 3:44 PM, Ethan A Merritt wrote:
>
>> On Tuesday, November 13, 2018 11:51:55 AM PST Zhijie Li wrote:
>> If somebody is going to send these
If somebody is going to send these files by email, please send one to me too.
Thanks in advance. I actually prefer to get a MTZ file because the miller
indices would serve as good clues for understanding the encodings. Even the
first 1024 bytes of an MTZ would do (data array starts at byte 80
Related by not exactly on topic: would anybody on the list be able to share old
map files (not MTZ:s) with Convex, Cray, Fujitsu, or VAX reals/strings? I’d be
interested to see what those files actually look(ed) like.
// Best wishes; Johan
> On Nov 9, 2018, at 18:38, Zhijie Li wrote:
>
> Hi
While working on PDB entries 1F0N and 1F0P, I aligned the side-chain
dipoles of 4 asparagines with the side-chain dipoles of tryptophans
(ring nitrogen is slightly negative, the rest of the 5-membered ring is
slightly positive). Aligning dipoles simultaneously optimized hydrogen
bonding for
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