CRYOCRYSTALLOGRAPHY: SUPPLIES & REFERENCES

Ken Goodwill & Duilio Cascio UCLA-DOE Laboratory

goodwill@mbi.ucla.edu, cascio@mbi.ucla.edu

Loop, pins, and magnets:

1)  Yale University Machine Shop  
John Squire  203 432-6300 fax=203432 6302 (or x3123 or x3059)
    - pins with loops  ~$9.50
    - pins without loops  ~$5.00
    - magnetic mounts  ~$5.00
2)  Hampton Research  Bob Cudney  714 699-1040
    - various thickness and diameter loops available.
      Thinnest loops recommended for least fiber diffraction

Cryo-tongs and other tools:  Note: Match the tongs to the pins!
1)  UC Berkeley Machine Shop
Eric Granlund  510 642-4486
    - cryo tongs  ~$225 each, 4 for $600
--> you may want to order one and then have your machine shop 
    make them 
2)  Hampton Research  Bob Cudney  714 699-1040

3)  McMaster-Carr Supply Co.  310 692-5911
    - very nice self-closing tweezers which once bent appropriately
    can be used to manipulate the pins by their bases, and which
    also form the basis for the UC Berkeley cryo tongs.
--> straight #7007A73 $4.77 each
--> curved   #7007A74 $4.77 each


Shipping and storage:

1)  GILMORE LIQUID AIR (Barry) 323-283 4721  
      DISTRIBUITOR of Taylor-Wharton   www.taylor-wharton.com
    - Taylor-Wharton cryogenic shipper, model CP100  $650 
    - molded shipping case for CP100  ~$280
    - long-term high-capacity storage Taylor-wharton 35VHC (holds 1260 vials) $1095
                                             Roller Base #R033-8C00           $ 165
    

2)  Fisher Scientific  800 766-7000

    - Corning cryogenic vials  2.0ml  #03-374-21  ~$100 for 500
    - Nalgene CryoCane holders  #15-350-20  ~$7 for 12
    - Nalgene CryosleevePVC    516-0001  VWR 24270-024
    - Fisher Low-Form Dewars for liquid nitrogen  # 10-196-6


References:
Petsko, G.A. (1975)  Protein crystallography at sub-zero temperatures:
 cryo-protective mother liquors for protein crystals.  
 J. Mol. Biol. 96, 381-392.

Hope, H. (1990)  Crystallography of biological macromolecules at ultra-low
 temperature.  Annual Rev. Biophys. Biophys. Chem. 19, 107-126

Tilton, R.F., Dewan, J.C., Petsko, G.A. (1992)  Effects of temperature
 on protein structure and dynamics:  X-ray crystallographic studies of the
 protein ribonuclease-A at nine different temperatures from 98-320K.
 Biochemistry 31, 2469-2481.

Rodgers, D.W. (1994)  Cryocrystallography.  Structure 2, 1135-1139.

Flash Freezing of Protein Crystals: Investigation of Mosaic Spread
and Diffraction Limit with Variation of Cryoprotectant Concentration.
E.P.Mitchell & E.F.Garman J.Applied Crystallogr. 27(6) 1070-1074 1Dec94

The effect of cryoprotectant(glycerol) upon the mosaicity and diffraction limit
of flash-frozen crystal of phosphorylase b crystals. Glycerol concentration was
varied between 0-70% v/v. Results show a well defined optimum in glycerol
concentration giving lowest mosaicity, maximum resolution, and no ice formation.
Demonstrating of systematic determination using minimum # of xtals.


 Glycerol concentrations required for cryoprotection of 50 typical protein
crystallization solutions. E.F. Garman & E.P. Mitchell. 
J. Appl. Cryst. 29, 584-587 (1996).
Table gives minimum glycerol concentrations for obtaining an amorphous glass
on freezing each of the Hampton Research Crystal Screen I solutions.