858-678-8765 
Fax: 413-751-0277 
purify@proteinx.com


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   ___ .