Home Protein Tools Lambda Protein Phosphatase (Lambda PP)

Lambda Protein Phosphatase (Lambda PP)
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Lambda Protein Phosphatase is a Mn2+-dependent protein phosphatase with activity towards phosphorylated serine, threonine and tyrosine residues. 

  • Dual specificity: can be used to release phosphate groups from phosphorylated serine, threonine and tyrosine residues in proteins
  • Recombinant enzyme with no detectable DNase, RNase or protease contaminating activities
  • Optimal activity and stability for up to 24 months
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Catalog # Cat # Concentration Conc Size List Price List Price Your Price Quantity
P0753S 400,000 units/ml 20,000 units $193.00 $193.00
P0753L 400,000 units/ml 100,000 units $769.00 $769.00
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  • Lambda Protein Phosphatase (Lambda PP) is a Mn2+-dependent protein phosphatase with activity towards phosphorylated serine, threonine and tyrosine residues. It is the 221 amino-acid product of the ORF221 open reading frame on bacteriophage lambda (1,2).

    Highlights

  • Dual Specificity: protein serine/threonine/tyrosine phosphatase 
  • Bacteriophage lambda, recombinant (E. coli)

    • Product Source

    Isolated from a strain of E. coli that carries the bacteriophage lambda ORF221 open reading frame under the control of a T7 expression system.
    This product is related to the following categories:
    Protein Phosphatases
    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
      • P0753S     -80    
          Lambda Protein Phosphatase (Lambda PP) P0753SVIAL -80 1 x 0.05 ml 400,000 units/ml
          NEBuffer Pack for Protein MetalloPhosphatases (PMP) B0761SVIAL -20 1 x 1 ml 10 X
          MnCl2 B1761SVIAL -20 1 x 1 ml 10 X
      • P0753L     -80    
          Lambda Protein Phosphatase (Lambda PP) P0753LVIAL -80 1 x 0.25 ml 400,000 units/ml
          NEBuffer Pack for Protein MetalloPhosphatases (PMP) B0761SVIAL -20 1 x 1 ml 10 X
          MnCl2 B1761SVIAL -20 1 x 1 ml 10 X

    • Properties & Usage

      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

    • Advantages and Features

      Application Features

      • Lambda PP can be used to release phosphate groups from phosphorylated serine, threonine and tyrosine residues in proteins. Note that different proteins are dephosphorylated at different rates.

    • Related Products

      Companion Products

      Materials Sold Separately

    • Product Notes

      1. 100 units of Lambda PP remove ~100% of phosphates (0.5 nmol) in phosphorylated myelin basic protein (phospho-MyBP, 18.5 kDa) in 30 minutes in a 50 µl reaction. The concentration of phospho-MyBP is 10 µM with respect to phosphate.
      2. The Protein Serine/threonine Phosphatase (PSP) activity of Lambda PP is assessed on MyBP phosphorylated exclusively on serine/threonine residues with cAMP-dependent Protein Kinase. The Protein Tyrosine Phosphatase (PTP) activity is assessed on MyBP phosphorylated exclusively on tyrosine residues with Abl Protein Tyrosine Kinase.
      3. Lambda PP is active on phosphorylated histidine residues (2).
      4. Optimal incubation times and enzyme concentrations must be determined empirically for each particular substrate.
      5. If the source of phosphorylated protein is a crude extract of cells or tissue, it is very important to include the appropriate protease inhibitors in the lysis buffer and to use shorter incubation time for dephosphorylation.
      6. The following levels of inhibition of Lambda PP (100 units) are found when the reaction buffer and MnCl2 are supplemented with: 
        • 10 mM Sodium Orthovanadate (3): 80% 
        • 10 mM Sodium Orthovanadate, 50 mM Sodium Fluoride: 90%
        • 50 mM EDTA: 95% 
        • 1% Triton X-100: no 
        • 0.4% Nonidet P-40: no 
        • 0.025% Tween 20: no 
        • 0.5 M NaCl: 5% 
        • ATP Mix (10 mM MgCl2, 0.1 mM ATP): no
        • Protease Inhibitor Cocktail*: 10% 

        *Pepstatin A, leupeptin and aprotinin, 10 µg/ml each, 0.5 mM PMSF and 1 mM benzamidine

    • References

      1. Cohen, P.T.W. and Cohen, P. (1989). Biochem. J.. 260, 931-934.
      2. Zhuo, S. et al. (1993). J. Biol. Chem.. 268, 17754-17761.
      3. Gordon, J.A. (1991). Methods Enzymol.. 201, 477-482.

  • Protocols, Manuals & Usage

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