Lambda Protein Phosphatase (Lambda PP)

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

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.

Companion Products

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

Materials Sold Separately

• NEBuffer Pack for Protein MetalloPhosphatases (PMP)

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

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.

1. Typical reaction protocol for lambda protein phosphatase

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

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 & Change Notifications

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]

• P0753S_L_v1

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]

• P0753S_v1_10013524
• P0753L_v1_10021702
• P0753S_v1_10027154
• P0753S_v1_10039492
• P0753L_v1_10035574
• P0753S_v1_10043585
• P0753S_v1_10052067
• P0753S_v1_10057712
• P0753L_v1_10057715
• P0753L_v1_10065469
• P0753S_v1_10065468
• P0753S_v1_10081134
• P0753L_v1_10081132
• P0753S_v1_10088262
• P0753S_v1_10098020
• P0753L_v1_10105461
• P0753S_v1_10109443
• P0753S_v1_10117444
• P0753L_v1_10133834
• P0753L_v1_10145287
• P0753S_v1_10140386
• P0753L_v1_10148381
• P0753S_v1_10157356
• P0753S_v1_10163650
• P0753L_v1_10167822
• P0753S_v1_10167820

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

• Lambda Protein Phosphatase (Lambda PP)
• NEBuffer Pack for Protein MetalloPhosphatases (PMP)
• MnCl₂

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