New England Biolabs, P0753S, Lambda Protein Phosphatase (Lambda PP)

Lambda Protein Phosphatase is a Mn Related Categories Protein Phosphatases Applications Protein Phosphatases and Kinases Specification Unit Definition One unit is defined as the amount of enzyme that hydrolyzes 1 nmol of p -Nitrophenyl Phosphate (50 mM) (NEB #P0757) in 1 minute at 30°C in a total reaction volume of 50 μl. Reaction Conditions 1X NEBuffer Pack for Protein MetalloPhosphatases (PMP) Supplement with 1 mM MnCl2 Incubate at 30°C 1X NEBuffer Pack for Protein MetalloPhosphatases (PMP) 50 mM HEPES 100 mM NaCl 2 mM DTT 0.01% Brij 35 (pH 7.5 @ 25°C) Storage Buffer 50 mM HEPES 100 mM NaCl 2 mM DTT 0.01% Brij 35 0.1 mM EGTA 0.1 mM MnCl2 50% Glycerol pH 7.5 @ 25°C Heat Inactivation 65°C for 60 minutes Molecular Weight Theoretical: 25 kDa Specific Activity 800,000 units/mg Storage Notes Avoid repeated freeze/thaw cycles. Shipping Notes Ships on dry ice FAQ Q: Can you recommend a cell lysis buffer that is compatible with Lambda Protein Phosphatase (NEB #P0753)? A: Lambda Protein Phosphatase (Lambda PP) is very sensitive to different reaction buffers. SDS at any concentration inhibits the enzyme. Ideally, the lysis buffer should have the closest composition to the optimal recommended buffer for Lambda PP, which is: 1X NEBuffer for PMP: 50 mM HEPES, 100 mM NaCl, 2 mM DTT, 0.01% Brij-35, pH 7.5, supplemented with 1mM MnCl2. These buffers are supplied with the enzyme. The ideal lysis buffer can also contain any mild non-ionic detergent such as 1% NP-40 or Triton X-100. If the PMP buffer is not ideal, another recommended lysis buffer is: 20 mM Tris-HCl, 150 mM NaCl, 1 mM EDTA, 1 mM EGTA, 1% Triton X-100, 1 µg/ml leupeptin, pH 7.5.