Home Protein Tools Products Casein Kinase II (CK2)

Casein Kinase II (CK2)
NEBU recombinant 30

Casein Kinase II (CKII) is a constitutively active serine/threonine protein kinase composed of two 44 kDa catalytic α-subunits and two 26 kDa regulatory β-subunits in an α2β2 configuration to form stable heterotetramers. The general recognition motif for phsophorylation by CKII is SXXE/D.

  • Recombinant enzyme with no detectable protease or phosphatase contaminating activities
  • Optimal activity and stability for up to 12 months
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Catalog # Concentration Size List Price Your Price Quantity
P6010S 500,000 units/ml 10,000 units $179.00
P6010L 500,000 units/ml 50,000 units $714.00
Please enter a quantity for at least one size
  • Casein Kinase II (CK2) is a constitutively active serine/threonine protein kinase composed of two 44 kDa catalytic α-subunits and two 26 kDa regulatory β-subunits in an α2β2 configuration to form stable heterotetramers. CK2 holoenzyme undergoes autophosphorylation at two serine residues (S2/S3) of its β-subunit. Recently it has been shown that CK2 α-subunits undergo intermolecular tyrosine-autophosphorylation at Y182, which may represent a specific regulatory mechanism. Also, CK2 is able to phosphorylate, under special circumstances, tyrosyl residues in proteins. CK2 is implicated in a variety of cellular functions (1,2).

    Product Source

    Isolated from a strain of E. coli expressing both α and β CK2 subunits derived from a human glioblastoma cDNA library (kindly provided by Dr. D. Marshak) (3).

    Recognition Determinant

    The CK2 substrate specificity is invariably determined by multiple acidic residues located at positions between -2 and +5 relative to the target amino acid (mostly Ser and rarely Thr). The general recognition motif for phsophorylation by CK2 is SXXE/D, although SXE/D and S/D, and variations of these sequences are also phosphorylated. Polyanionic compounds, like heparin, inhibit CK2 activity with a Ki of 1.4 nm (4,5).
    This product is related to the following categories:
    Protein Kinases Products
    This product can be used in the following applications:
    Protein Phosphatases and Kinases

    • Reagents Supplied

      The following reagents are supplied with this product:

      NEB # Component Name Component # Stored at (°C) Amount Concentration
      • P6010S     -80    
          Casein Kinase II (CK2) P6010SVIAL -80 1 x 0.02 ml 500,000 units/ml
          NEBuffer™ for Protein Kinases (PK) B6022SVIAL -20 1 x 1 ml 10 X
      • P6010L     -80    
          Casein Kinase II (CK2) P6010LVIAL -80 1 x 0.1 ml 500,000 units/ml
          NEBuffer™ for Protein Kinases (PK) B6022SVIAL -20 1 x 1 ml 10 X

    • Properties & Usage

      Unit Definition

      One unit is defined as the amount of CK2 required to catalyze the transfer of 1 pmol of phosphate to CK2 Peptide Substrate, RRRADDSDDDDD (100 µM), in 1 minute at 30°C in a total reaction volume of 25 µl (4,5).

      Reaction Conditions

      1X NEBuffer™ for Protein Kinases (PK)
      Supplement with 200 µM ATP
      Incubate at 30°C

      1X NEBuffer™ for Protein Kinases (PK)
      50 mM Tris-HCl
      10 mM MgCl2
      0.1 mM EDTA
      2 mM DTT
      0.01% Brij 35
      (pH 7.5 @ 25°C)

      Storage Notes

      • Avoid repeated freeze/thaw cycles.

    • Related Products

      Companion Products

      Materials Sold Separately

    • Product Notes

      1. Molecular Weight: α-subunit (45 kDa), β-subunit (25 kDa). The apparent molecular weight of the α-subunit estimated by SDS-PAGE is about 42 kDa.
      2. General notes:
        • For short term storage (two weeks or less) CK2 can be stored at -20°C. 
        • If the source of protein to be phosphorylated is a crude extract of cells or tissue, it is very important to include the appropriate protease and protein phosphatase inhibitors in the lysis buffer and to use shorter incubation time for phosphorylation. 
        • Reaction Conditions: 1X NEBuffer for Protein Kinases (PK), supplement with 200 µM ATP and gamma-labeled ATP to a final specific activity of 100-500 µCi/µmol. (NEBuffer for Protein Kinases (PK) will also accept GTP as a phosphoryl donor in place of ATP).
      3. Usage notes:
        • Optimal incubation times and enzyme concentrations must be determined empirically for each particular substrate. 
        • If possible, the ATP concentration should be at or near saturation (5 -10-fold over Km). Apparent Km values of ATP for most protein kinases are below 100 μM. 
        However, if the objective is to measure enzyme activity using gamma-labeled ATP, it is best to use 100-200 μM ATP in order to have higher specific activity of gamma-labeled ATP (100-500 cpm/pmol). Also, an excess of substrate should be used, and the level of phosphorylation should not exceed 10% for determination of the initial rate.

        To phosphorylate a protein or peptide substrate to completion, the ATP concentration should be about 5-fold over the limited substrate concentration. Higher enzyme concentration and prolonged incubation times should be employed.

        Recommended reference:Protein Phosphorylation: A Practical Approach (1993) ed. Hardie, D.G. IRL Press.

    • References

      1. Chester, N. and Marshak, D.R. (1993). Anal. Biochem. 209, 284-290.
      2. Donella-Deana, A. et al. (2001). Biochem J. 357, 563-567.
      3. Marin, O. et al. (1999). J. Biol. Chem. 274, 29260-29265.
      4. Marin, O. et al. (1994). BBRC. 198, 898-905.
      5. Sarno, S. et al. (1996). J. Biol. Chem. 271, 10595-10601.

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