Protein crystallography prescreen kit

• US 20050205006A1
• Filed: 03/17/2005
• Published: 09/22/2005
• Est. Priority Date: 08/14/2002
• Status: Active Grant

• Current Assignee: Lawrence Livermore National Security LLC (Government of the United States of America)
• Original Assignee: Regents of the University of California (University of California)
• Inventors: Segelke, Brent W., Rupp, Bernhard, Krupka, Heike I.
• Granted Patent: US 7,276,216 B2
• Time in Patent Office: Days
• US Class Current: 117/206
• CPC Class Codes: B01L 1/52 Transportable laboratories; Field kits
• CPC Class Codes: C30B 29/58 Macromolecular compounds
• CPC Class Codes: C30B 7/00 Single-crystal growth from solutions using solvents which are liquid at normal temperature, e.g. aqueous solutions from molten solvents C30B9/00; by normal or gradient freezing C30B11/00; under a protective fluid C30B27/00
• CPC Class Codes: G01N 2001/4027 evaporation leaving a concentrated sample
• CPC Class Codes: Y10S 117/901 Levitation, reduced gravity, microgravity, space
• CPC Class Codes: Y10T 117/1024 for crystallization from liquid or supercritical state

Abstract

A kit for prescreening protein concentration for crystallization includes a multiplicity of vials, a multiplicity of pre-selected reagents, and a multiplicity of sample plates. The reagents and a corresponding multiplicity of samples of the protein in solutions of varying concentrations are placed on sample plates. The sample plates containing the reagents and samples are incubated. After incubation the sample plates are examined to determine which of the sample concentrations are too low and which the sample concentrations are too high. The sample concentrations that are optimal for protein crystallization are selected and used.

Claims

1. A kit for prescreening protein concentration for crystallization, comprising:

a multiplicity of vials,

a multiplicity of reagents,

a multiplicity of sample plates, and

instructions for using said vials, said reagents, and said sample plates for assessing the proper protein concentration to carry out protein crystallization.

2. The kit for prescreening protein concentration for crystallization of claim 1, wherein said multiplicity of vials are cryo-vials.

3. The kit for prescreening protein concentration for crystallization of claim 2, wherein said multiplicity of cryo-vials are Eppendorf cryo-vials.

4. The kit for prescreening protein concentration for crystallization of claim 3;

wherein said multiplicity of Eppendorf cryo-vials are 1.5 ml. Eppendorf cryo-vials.

5. The kit for prescreening protein concentration for crystallization of claim 1, wherein said vials are labeled.

6. The kit for prescreening protein concentration for crystallization of claim 5, wherein said vials are labeled with cryo-babies labels.

7. The kit for prescreening protein concentration for crystallization of claim 1, wherein said multiplicity of reagents are precisely formulated and consist of high quality chemicals of per analysis grade chemicals.

8. The kit for prescreening protein concentration for crystallization of claim 7, wherein said multiplicity of reagents are aqueous solutions containing 0.1M Tris/HCl buffer at pH 8.5.

9. The kit for prescreening protein concentration for crystallization of claim 1, wherein said multiplicity of reagents includes a reagent of 50% PEG 4000.

10. The kit for prescreening protein concentration for crystallization of claim 1, wherein said multiplicity of reagents includes a reagent of 30% PEG 4000.

11. The kit for prescreening protein concentration for crystallization of claim 1, wherein said multiplicity of reagents includes a reagent of 5% PEG 4000.

12. The kit for prescreening protein concentration for crystallization of claim 1, wherein said multiplicity of reagents includes a reagent of 3M Ammonium Sulfate.

13. The kit for prescreening protein concentration for crystallization of claim 1, wherein said multiplicity of reagents includes a reagent of 2M Ammonium Sulfat.

14. The kit for prescreening protein concentration for crystallization of claim 1, wherein said multiplicity of reagents includes a reagent of 0.5M Ammonium Sulfate.

15. The kit for prescreening protein concentration for crystallization of claim 1, wherein said multiplicity of reagents includes a reagent of 40% Isopropanol.

16. The kit for prescreening protein concentration for crystallization of claim 1, wherein said multiplicity of reagents includes a reagent of 30% Isopropanol.

17. The kit for prescreening protein concentration for crystallization of claim 1, wherein said multiplicity of reagents includes a reagent of 5% Isopropanol.

18. The kit for prescreening protein concentration for crystallization of claim 1, wherein said multiplicity of reagents include a first reagent of 50% PEG 4000, a second reagent of 30% PEG 4000, a third reagent of 5% PEG 4000, a fourth reagent of 3M Ammonium Sulfate, a fifth reagent of 2M Ammonium Sulfate, a sixth reagent of 0.5M Ammonium Sulfate, a seventh reagent of 40% Isopropanol, an eight reagent of 30% Isopropanol, and a ninth reagent of 5% Isopropanol.

19. The kit for prescreening protein concentration for crystallization of claim 1, wherein said instructions include directions for mixing reagents with a protein sample.

20. The kit for prescreening protein concentration for crystallization of claim 19, wherein said instructions include directions to mix 1 L1 of said reagents with lAl of said protein sample in drops of various concentrations on said plates.

21. The kit for prescreening protein concentration for crystallization of claim 20, wherein said instructions include directions to control said drops after 1-5 min incubation time under a light microscope.

22. The kit for prescreening protein concentration for crystallization of claim 1, wherein said instructions indicate that if all reagents produce clear drops the protein concentration is too low, and if all reagents produce heavy precipitant, turning the drop opaque, the protein concentration is too high for protein crystallization to occur.

23. The kit for prescreening protein concentration for crystallization of claim 1, wherein said multiplicity of vials, said multiplicity of reagents, said multiplicity of sample plates, and said instructions are packed in a container.