Lambda Protein Phosphatase

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SKU: 02-300 Research Area: Applications:

Description

λ Protein Phosphatase (λ-PPase) is a Mn 2-dependent protein phosphatase with activity towards phosphorylated serine, threonine, tyrosine and histidine residues.  It is the 221 amino-acid product of ORF221 open reading frame on bacteriophage lambda (1, 2).  λ-PPase was expressed as a recombinant protein in E. coli and highly purified (2).  This product is an intact enzyme of high quality without tag.

 

Applications: 

λ-PPase can be used to release phosphate groups from phosphorylated serine, threonine, tyrosine and histidine residues in proteins (2).  It should be noted that different proteins are dephosphorylated at different rates.  Optimal reaction temperature is 30°C.  Inclusion of protease inhibitor cocktail and shortest incubation time is desired when assays are done with crude samples.

 

Form: 400 U/ul λ-PPase in 50mM HEPES (pH 7.5), 100mM NaCl, 2mM dithiothreitol, 0.1 mM MnCl2,  0.1mM EDTA, 50% glycerol, 0.01% Brij 35.

 

Storage: Store at -20°C for a week (for longer period, at -70°C and avoid repeating of freezing- thawing)

 

Activity:

400 U/ul, where one unit is defined as the amount of enzyme that hydrolyzes 1 nmole of

p-nitrophenyl phosphate per minute at 30°C. Unit definition assays are performed with 50mM

p-nitrophenyl phosphate in λ-PPase buffer, supplemented with 2 mM MnCl2 in a 50 ul reaction.

 

Specific Activity: 400,000 U/mg

 

Quality Assurance: Greater than 95% homogeneous protein determined by SDS-PAGE (CBB staining) that contains no detectable protease activity

 

Reagents Supplied with Enzyme:

10 x λ-PPase Reaction Buffer [500mM Tris-HCl (pH 7.6), 1M NaCl, 20mM  dithiothreitol, 1mM EDTA, 0.1% Brij 35]

10 x Mn2 (20 mM MnCl2)

 

Data Link: UniProtKB/Swiss-Prot P03772 (PP_LAMBD)

 

References:

  1. Cohen PTW & Cohen P (1989) “Discovery of a protein phosphatase activity encoded in the genome of bacteriophage λ.” Biochem J. 260: 931-934 PMID: 2548489

 

  1. Zhuo S et al (1993) “Expression, purification, crystallization, and biochemical characterization of recombinant protein phosphatase.” J. Biol. Chem.268:17754-17761 PMID: 8394350

 

Fig. 1 SDS-PAGE of in λ-PPase

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02-300_lambda protein phosphatase_03222018 application/pdf 158 KB

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