Name: __________________
Phone: __________________
Address: __________________
Fax: __________________
__________________ Email: __________________
__________________
About your problematic protein
1. Has the construct been sequenced?
Yes ___ , No ___ .
2. What kind of construct it is?
Home-made _____________ , Commercial
_________ (please specify it).
3. The Molecular Weight of the protein:
Itself _______ kd, Fusion protein _______
kd.
4. What is the origin of the protein?
Prokaryotic _______ , Eukaryotic ________ .
5. What is the problem with the protein?
Expression
Protein is expressed, but degraded. ___
Protein is expressed as "inclusion body". ___
Protein does not express, or expression level is below 1mg/L.
___
Protein is toxic, cell die after induction. ___
Purification
You have tried: Affinity chromatography
___
Ion-exchange chromatography ___
Gel filtration chromatography ___
Reverse phase chromatography ___
HIC chromatography ____
Hydroxyapatite chromatography ___
Bulk separation: Precipitation (i.e. Ammonium Sulfate, organic) ____
,
Adsorption ___
Ultra filtration ___ .
Solubility
Protein is not soluble ___
Protein is precipitated out of solution in the process of purification
___
Solublization of protein has been tried by using:
GuHCl ___
Urea ___
High/Low pH ___
Organic solvent ___
Refolding has been tried at the condition: ________
Activity
After purification, protein is inactive ___
After refolding, protein has no activity ___
After storage for a while, protein lost the activity ___
How do you measure the activity? ___
The protein sample you need
6. How much protein do you need?
2~3mg ___ , 20~30mg
___ , 200~300mg ___ .
7. How pure protein do you need?
Affinity elute ___
FPLC pure ___
Electrophoresis pure ___
Clinical pure ___
8. What kind of sample do you prefer?
Powder ___ , Solution ___ .
9. What protein concentration do you prefer?
0.1mg/ml ___ , 1mg/ml ___ ,
10mg/ml ___ .
10. What buffer system do you prefer? Please specify.
_____________
11. What additives do you prefer (yes)? or want to avoid (no)? Please
specify.
EDTA/EGTA ___
Divalent ion ___
Reductant ___
Protease inhibitor ___
Detergent ___
Stabilizing agent: Salt
____ , Glycerol/sugar ___ ,
Other ___ .
Additional info
12. Are you interested in the kinetic measurement of protein binding
to other biomolecule through Biacore?
Yes ___ , No ___
13. Are you interested in the structure determination through crystallization
or NMR?
Yes ___ , No ___
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