Lambda Protein Phosphatase (Lambda PP) |NEB酶试剂 New England Biolabs

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产品信息

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).

重点

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

    • 产品来源

    Isolated from a strain of E. coli that carries the bacteriophage lambda ORF221 open reading frame under the control of a T7 expression system.

    产品类别:
    Protein Phosphatases Products

    应用:
    Protein Phosphatases and Kinases

    • 产品组分信息

      本产品提供以下试剂或组分：

      NEB # 名称 组分货号 储存温度 数量 浓度
      • 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

    • 特性和用法

      单位定义

      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.

      反应条件

      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)

      贮存溶液

      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

      热失活

      65°C for 60 minutes

      分子量

      理论上的: 25 kDa

      比活度

      800,000 units/mg

      存储注意事项

      • Avoid repeated freeze/thaw cycles.

      货运单

      • Ships on dry ice

    • 优势和特性

      应用特性

      • 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.

    • 相关产品

      相关产品

      • p-Nitrophenyl Phosphate (PNPP)
      • Sodium Orthovanadate (Vanadate)

      单独销售的组分

      • NEBuffer Pack for Protein MetalloPhosphatases (PMP)

    • 注意事项

      1. The following information can be used as suggested initial conditions for dephosphorylation of proteins with Lambda PP.
      2. 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.
      3. 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.
      4. Lambda PP is active on phosphorylated histidine residues (2).
      5. 0ptimal incubation times and enzyme concentrations must be determined empirically for each particular substrate. 
      6. 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.
      7. 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

    • 参考文献

      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.

    操作说明、说明书 & 用法

    • 操作说明

      1. Typical reaction protocol for lambda protein phosphatase

    FAQs & 问题解决指南

    • FAQs

      1. Can you recommend a cell lysis buffer that is compatible with Lambda Protein Phosphatase (NEB #P0753)?