New England Biolabs, P6010S, Casein Kinase II (CK2)

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. Related Categories Protein Kinases Applications Protein Phosphatases and Kinases Specification Unit Definition One unit of enzyme catalyzes the phosphorylation of 1 pmol of RRRDDD S DDD substrate in 10 minutes at 30˚C. 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 EGTA 2 mM DTT 0.01% Brij 35 (pH 7.5 @ 25°C) Storage Buffer 20 mM Tris-HCl 350 mM NaCl 1 mM EDTA 2 mM DTT 0.1% Triton® X-100 pH 7.5 @ 25°C Storage Notes Avoid repeated freeze/thaw cycles. FAQ Q: Can you recommend inhibitors for CK2 (P6010)? A: Heparin inhibits at a Ki of 1.4 nM. Q: How much Casein Kinase II (NEB# P6010) should be used? A: The amount of enzyme required varies when different substrates are used. Start with 1-2 µl for 1µg of protein for one hour in a 10-25 µl reaction. If there is still unphosphorylated material, let the reaction go overnight. Q: What is the consensus sequence for this CK2 (P6010)? A: The consensus sequence is SXXE/D. This kinase phosphorylates serine or threonine residues that are N-terminal to acidic amino acids.