Materials and Reagents

1. Escherichia coli (E.coli) strains - for cloning and for expression - DH5α, Rosetta (DE3 pLysS)
   
2. Specific synthetic oligonucleotides
   
3. PCR reagents (dNTP, MgCl₂, PCR buffer, Taq-polymerase, DNA) (Thermo Fisher Scientific, catalog numbers: R0181 , EP0612 )
   
4. Plasmids for cloning and for expression [pTZ57R/T, pET41a(+)]
   
5. CaCl₂ (Sigma-Aldrich, catalog number: C3306-250G ) (for transformation)
   
6. Antibiotics (Ampicillin, Kanamycine, Chloramphenicol) (AppliChem, catalog numbers: A-0839 , A-1493 , A-6435 )
   
7. Isopropyl thio-β-D-galactoside (IPTG) (Fermentas, catalog number: R0393 )
   
8. Tris-HCl (Sigma-Aldrich, catalog number: T6791 )
   
9. Ethylenediaminetetraacetic acid (EDTA) (AppliChem, catalog number: A-1104.0500 )
   
10. Dithiothreitol (DTT) (Sigma-Aldrich, catalog number: 43815-5G )
    
11. Ethylene glycol-bis(2-aminoethylether)-N,N,N′,N′-tetraacetic acid (EGTA) (Sigma-Aldrich, catalog number: E3889-10G )
    
12. Na₃VO₄ (Sigma-Aldrich, catalog number: S6508-10G )
    
13. NaF (Sigma-Aldrich, catalog number: S7920-100G )
    
14. β-glycerophosphate (Sigma-Aldrich, catalog number: G9422-50G )
    
15. Sucrose (AppliChem, catalog number: A-1125 )
    
16. Phenylmethylsulfonyl fluoride (PMSF) (AppliChem, catalog number: A-0999 )
    
17. Benzamidine hydrochloride (Sigma-Aldrich, catalog number: B6506 )
    
18. 6-aminocaproic acid (Sigma-Aldrich, catalog number: A7824-25G )
    
19. 2-(N-morpholino)ethanesulfonic acid (MES) (Amresco, catalog number: Am-E169-0.05 )
    
20. Myelin Basic Protein bovine (MBP) (Sigma-Aldrich, catalog number: M1891-5MG )
    
21. Histone from calf thymus (H1) (Sigma-Aldrich, catalog number: H5505-100MG )
    
22. Bovine serum albumin (BSA) (Sigma-Aldrich, catalog number: 05470-1G )
    
23. MgCl₂ (AppliChem, catalog number: A-1447 )
    
24. Brilliant Blue R Concentrate (Sigma-Aldrich, catalog number: B8647 )
    
25. BCA Protein Assay Kit (Pierce Thermo Scientific, catalog number: 23227 )
    
26. Sodium dodecyl sulfate, SDS (Sigma-Aldrich, catalog number: L3771 )
    
27. TGX FastCast Acrylamide Starter Kit (12%) (Bio-Rad Laboratories, catalog number: 161-0175 )
    
28. N,N,N',N'-tetramethylethylenediamine (TEMED) (Bio-Rad Laboratories, catalog number: 161-0800 )
    
29. Ammonium Persulfate (APS) (Bio-Rad Laboratories, catalog number: 161-0700 )
    
30. Illustra NAP-5 Columns (GE Healthcare Life Sciences, catalog number: 17-0853-02 )
    
31. [γ-³²P] ATP
    
32. Homogenization buffer (see Recipes)
    
33. Reaction mixture (see Recipes)
    

Equipment

1. Temperature controlled shaker (Heidolph, model: Unimax 1010 )
   
2. French Pressure Cell Press (Aminco) or glass beads (Sigma-Aldrich, catalog number: G4649 ) for cell disruption
   
3. Thermostat
   
4. Spectrophotometer
   
5. Gel electrophoresis equipment (Mini-PROTEAN Tetra Handcast Systems, Bio-Rad Laboratories, catalog number: 165-8000 )
   
6. Laminar flow box
   
7. Refrigerated microcentrifuge preset to 4 °C (Eppendorf, model: 5415 R )
   
8. Gel Dryer System (Bio-Rad Laboratories, catalog number: 165-1789 )
   
9. Exposure cassette/BioMax^® intensifying screen set (Sigma-Aldrich, catalog numbers: C4979 and P8432 )
   
10. Carestream^® Kodak^® autoradiography GBX developer/replenisher (Sigma-Aldrich, catalog number: P7042 )
    
11. Carestream^® Kodak^® autoradiography GBX fixer/replenisher (Sigma-Aldrich, catalog number: P7167 )
    

Procedure