data for structural and crystallization communications - example

Example 4: microbatch or liquid-liquid diffusion or dialysis method

Crystallization 4: Crystals of a protein were obtained by the microbatch method using Nunc HLA plates (Nalge Nunc International). Each crystallization drop was prepared by mixing 1.0 μl precipitant solution (12.5% PEG 4000, 0.1 M MES, pH 5.9) and 1.0μl protein solution (25 mg/ml in 20 mM Tris HCl, 0.2 M NaCl, pH 8.0). The crystallization drop was overlaid with a 1:1 mixture of silicone/paraffin oils, allowing slow evaporation of the drop at 296 K.
Cryo-conditions for crystal 4: Prior to data collection, single crystals were soaked in three different cryoprotectant solutions for 1 min each and then transferred to liquid nitrogen. The cryoprotectant solutions contained reservoir solution supplemented with 5, 20, and 25% glycerol.
_exptl_crystal.crystal_id              4
_exptl_crystal.size_max                ?
_exptl_crystal.size_mid                ?
_exptl_crystal.size_min                ?

_exptl_crystal_grow.crystal_id         4
_exptl_crystal_grow.method             'microbatch'
_exptl_crystal_grow.method_details
; Droplets were in Nunc HLA plates and covered with a 1:1 mixture of
  silicone and paraffin oils, which allowed slow evaporation.
;
_exptl_crystal_grow.temp               296

loop_
_pdbx_exptl_crystal_grow_sol.crystal_id
_pdbx_exptl_crystal_grow_sol.sol_id
_pdbx_exptl_crystal_grow_sol.volume
_pdbx_exptl_crystal_grow_sol.volume_units
_pdbx_exptl_crystal_grow_sol.pH

4    'protein'      1    'microlitre'    8.0
4    'precipitant'  1    'microlitre'    5.9

loop_
_pdbx_exptl_crystal_grow_comp.crystal_id
_pdbx_exptl_crystal_grow_comp.sol_id
_pdbx_exptl_crystal_grow_comp.comp_id
_pdbx_exptl_crystal_grow_comp.comp_name
_pdbx_exptl_crystal_grow_comp.conc
_pdbx_exptl_crystal_grow_comp.conc_range
_pdbx_exptl_crystal_grow_comp.conc_units

4  'protein'       1   'protein'     25.   .    'mg/ml'
4  'protein'       2   'Tris HCl'    20.   .    'millimolar'
4  'protein'       3   'NaCl'        0.2   .    'molar'

4  'precipitant'   1   'PEG 4000'    12.5  .    'percent_weight_by_volume'
4  'precipitant'   2   'MES'         0.1   .    'molar'

_pdbx_exptl_crystal_cryo_treatment.crystal_id  4

_pdbx_exptl_crystal_cryo_treatment.final_solution_details
; 25% (v/v) glycerol in precipitant solution
;

_pdbx_exptl_crystal_cryo_treatment.soaking_details
; A series of 1 min soaks beginning at 5% (v/v) glycerol in precipitant,
  transiting to 20% (v/v) glycerol, and finally to 25% glycerol
;

_pdbx_exptl_crystal_cryo_treatment.cooling_details
; Direct immersion in liquid nitrogen.
;

_pdbx_exptl_crystal_cryo_treatment.annealing_details
; None.
;


How this example will appear in the journal

Crystallization
Crystallization method Microbatch  
Temperature 296
Additional details Droplets were in Nunc HLA plates and covered with a 1:1 mixture of silicone and paraffin oils, which allowed slow evaporation.
Crystallization solutions
   Protein (1 μl) Protein 25 mg ml-1, Tris 20 mM, NaCl 0.2 M, pH 8.0
   Precipitant (1 μl) PEG 4000 12.5% (w/v), MES 0.1 M pH 5.9  
Cryo treatment
  Final cryoprotection solution 25% (v/v) glycerol in precipitant solution
  Soaking A series of 1 min soaks beginning at 5% (v/v) glycerol in precipitant, transiting to 20% (v/v) glycerol, and finally to 25% glycerol
  Cooling Direct immersion in liquid nitrogen
  Annealing None



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