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the University of Glasgow.
Please find full details listed here (Referen
Dear all
Do you enjoy purifying challenging proteins? Do you want to support antibody
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Then consider applying for these roles in our Protein Production group:
Senior Scientist - Protein Biochemist job in Slo
Hi Amala,
1. Did you verify the sequence or the presence of the His-tag? If you were
not the person for cloning. I used to spend 14 months working on an clone
and eventually I was allowed to check the sequence and verify the
expression of His-tag by Western-Blot. There was no his-tag.
2. You can t
Dear Amala
Please increase NaCl concentration to 200mM from 50mM. That can help you
out by increasing affinity of your protein to bead and will delay the
elution time.
On Mon, Aug 13, 2018, 7:20 PM amala mathimaran wrote:
> Dear All
>
> I am working with HIS – tag protein in N-terminal (hexa hi
Hello Amala,
Usually Ni-NTA won't have this kind of problem of binding. Probably your
protein is have interaction with hexagon his tag which is affecting its
affinity towards beads. You can try putting tag in C- terminal end of the
protein.
On Mon, 13 Aug 2018, 8:02 pm Artem Evdokimov,
wrote:
Hi Amala,
Depending on the resin you used there may be a conflict with BME and also
50 mM TRIS may be too much at the pH 7.5. Easy to test: use HEPES pH 7.5
and TCEP instead of BME, or use 30 mM TRIS at pH 8.0
Alternatively your protein is aggregated and does not bind well...
Artem
- Cosmic Cat
Dear All
I am working with HIS – tag protein in N-terminal (hexa histidine). The
protein from Prokaryotic origin cloned into pET30a+ vector and
expressed in *E.coli
*BL21 cells. The expression was good. I am trying to purify a protein using
Ni-affinity column equilibrate with Buffer A 50mM Tris pH
Sajid,
As Antonio said, we need more information in order to really help you.
Particularly what columns you used and a little more about the protein target.
As one who has worked on mammalian tissues, I found that some proteins are very
sensitive to proteolysis after homogenization, but other
5de-dmarc-requ...@jiscmail.ac.uk]
Sent: 18 January 2017 11:32
To: CCP4BB@JISCMAIL.AC.UK
Subject: [ccp4bb] Protein Purification
Dear All
I am purifying a protein from liver. In first step I spotted protein in Flow
Through and could see intense band in SDS PAGE. I pooled the fraction and did
second
Dear All
I am purifying a protein from liver. In first step I spotted protein in Flow
Through and could see intense band in SDS PAGE. I pooled the fraction and did
second column. Surprisingly I could not see UV absorbance or even any band in
the SDS. I used PMSF as protease inhibitor from begin
An opening is available in the laboratory of Dr. Stephen Fesik
(http://www.mc.vanderbilt.edu/fesiklab) for the Research Associate position in
structural biology and drug discovery. The candidate must possess a degree in
biochemistry or a related field with 1-2 years of experience in protein
pur
Dear All,
1st of all , Thanks for the very quick feedback. I will answer your questions
to make the picture more clear-
Nicolas-
Yes the dimer is co-expressed. Almost all the protein is in the sup and not in
the pellet. Multiple-step dialysis has failed. I have been trying various
controls a
Do you really need to remove the NaCl? Some ionic strength is often
necessary to stabilize proteins. Our routine purification buffers all
contain at least 100 mM NaCl. This will not usually interfere with
crystallization screening.
To minimize the probability of aggregation, you need to (1) en
ur protein (
for example with Circular dichroisme or DLS).
Hope to help you.
Nicolas
De : CCP4 bulletin board [CCP4BB@JISCMAIL.AC.UK] de la part de Anindito Sen
[andysen.to...@gmail.com]
Envoyé : jeudi 30 janvier 2014 14:17
À : CCP4BB@JISCMAIL.AC.UK
Dear All,
This may be slightly off-the-track question but your feedback will be very much
appreciated. The situation is-
I obtain a very low amount of the protein of my interest (a hetro-dimer) from
the construct I am using (only 8% of the total amount of protein obtained is
the protein of my
---POSITIONS IN PROTEIN
PURIFICATION-
We seek an experienced individual with expertise in protein expression and
purification from prokaryotic and eukaryotic sources. This position resides
within the Macromolecular Therapeutics Development Facility (MTDF
Scientist/Senior Scientist position Protein purification, Evotec,
Abingdon, UK
Applications are sought for a Scientist/Senior Scientist position in the
Protein Production team at Evotec
Job Description:
Based at Abingdon, Oxfordshire, Evotec offer integrated services which
cover the entir
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