Another possibility you might like to consider is to make use of
non-covalent-binding dyes:
1) If your protein contains a His6-tag, you could add a bit of a (for
partial non-covalent labeling) Tris–nitrilotriacetic acid
(Tris-NTA)–conjugated based dye/fluorophore such as
Tris-NTA-Orange-Green.... See https://doi.org/10.2144/000113551
<https://doi.org/10.2144/000113551>
2) If no His6-tag, dependent on the protein nature (hydrophobic vs.
charged/hydrophylic, etc.), you could add (again partial labelling) make
use of cyanine or squarylium dyes... See https://doi.org/10.3390/90300040
- Jeroen -
Am 19.02.22 um 09:18 schrieb Casper Wilkens:
Is absorbance at 205 nm not a possibility, Jack?
Casper Wilkens
Asst. Prof.
Structural Enzymology & Biorefining
DTU Bioengineering
Technical University of Denmark
Department of Biotechnology and Biomedicine
Søltofts Plads
Building 224
Room 028
2800 Kgs. Lyngby
c...@dtu.dk <mailto:c...@bio.dtu.dk>
www.bioengineering.dtu.dk/ <http://www.bioengineering.dtu.dk/>
https://www.cazypedia.org/index.php/User:Casper_Wilkens
<https://www.cazypedia.org/index.php/User:Casper_Wilkens>
https://twitter.com/protein_artist <https://twitter.com/protein_artist>
https://www.researchgate.net/profile/Casper_Wilkens
<https://www.researchgate.net/profile/Casper_Wilkens>
------------------------------------------------------------------------
*Fra:* CCP4 bulletin board <CCP4BB@JISCMAIL.AC.UK> på vegne af Artem
Evdokimov <artem.evdoki...@gmail.com>
*Sendt:* 19. februar 2022 02:12:50
*Til:* CCP4BB@JISCMAIL.AC.UK
*Emne:* Re: [ccp4bb] Strategies for increasing Trp content of proteins
We have had success placing single trp on the outside of proteins, as
weird as it sounds it works pretty well. If your protein is easy to
express you can try a few choices.
If your protein is difficult to express, a fusion with a trp-positive
domain or with gfp may be a better option.
There are many other options but their application is best considered
with more information in hand
Artem
On Fri, Feb 18, 2022, 8:02 PM Tanner, John J. <tanne...@missouri.edu>
wrote:
Dear CCP4BB,
We are working on a protein that has no Trp residues, which makes
chromatography challenging due to the low absorbance at 280 nm
(Abs 0.1% = 0.104). Does anyone have experience using mutagenesis
to increase the Trp content of proteins?
Thanks,
Jack Tanner
--
John J. Tanner
Professor of Biochemistry and Chemistry
Associate Chair of Biochemistry
Department of Biochemistry
University of Missouri
117 Schweitzer Hall
503 S College Avenue
Columbia, MO 65211
Phone: 573-884-1280
Email: tanne...@missouri.edu <mailto:tanne...@missouri.edu>
https://cafnrfaculty.missouri.edu/tannerlab/
<https://cafnrfaculty.missouri.edu/tannerlab/>
Lab: Schlundt Annex rooms 3,6,9, 203B, 203C
Office: Schlundt Annex 203A
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signature.html *Dr. /math. et dis. nat./ Jeroen R. Mesters
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