Product Class: Other

Lambda Protein Phosphatase (Lambda PP)
NEBU cloned at NEB recombinant 30 65 Heat

Product Introduction

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
Catalog # Size Concentration
P0753S 20000.0 units 400000 units/ml
P0753L 100000.0 units 400000 units/ml

Product Information

Description

Highlights

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

    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 Products
    This product can be used in the following applications:
    Protein Phosphatases and Kinases

    Reagents Supplied

    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

    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.

    Product Notes

    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

    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

    Protocols

    1. Typical reaction protocol for lambda protein phosphatase

    FAQs & Troubleshooting

    FAQs

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

    Quality, Safety & Legal

    Quality Assurance Statement

    Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.

    Specifications

    The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]

    Certificate Of Analysis

    The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]

    Safety DataSheets

    The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

    Legal and Disclaimers

    Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email busdev@neb.com.

    This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

    New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.

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