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Lamda Protein Phosphatase (5 x 400 units/μl)

Product#: FNK-02-300-5

$441.75

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Lamda Protein Phosphatase (5 x 400 units/μl)

Cat No: FNK-02-300-5
Size: 5 x 20000 units (400U/ul)
Shipping and Storage: Sent at 4℃ or -20℃ and store at -20℃. Do not freeze below -20℃

Description

Lambda Protein Phosphatase ( l-PPase) 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).

Functional full-length recombinant λProtein Phosphatase (λ-PPase) expressed in E.coli. It 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). 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℃. 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.

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℃. 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

Purity

Greater than 95% as 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)

Fig.2 Dephosphorylation of phospho -NPM1 protein by incubation with λprotein phosphatase in vitro. HeLa cells treated with Nocodazole was lysed, and NPM1 protein was immunoprecipited with anti-NPM1 antibody. The precipitate was suspended in 50 ul of λ protein phosphatase reaction buffer added with 5 ul of the protein phosphatase and incubated at 30℃ for the indicated time (min). The reaction products were analyzed by western blotting.

Reference

This product has been used in the following publications.

Tomii S et al. Cortical Actin Alteration at the Matrix-Side Cytoplasm in Lung Adenocarcinoma Cells and Its Significance in Invasion. Pathobiology.2017; 84(4):171-183. PMID: 28002815
Tojima T et al. Steering neuronal growth cones by shifting the imbalance between exocytosis and endocytosis. J Neurosci. 2014 May 21;34(21):7165-78. PMID: 24849351
Moriyama T et al.SUMO-modification and elimination of the active DNA demethylation enzyme TDG in cultured human cells. Biochem Biophys Res Commun. 2014 May 9;447(3):419-24. PMID: PMID: 24727457

Useful References

Cohen PTW & Cohen P (1989) “Discovery of a protein phosphatase activity encoded in the genome of bacteriophage ?.” Biochem J. 260: 931-934 PMID: 2548489
Zhuo S et al (1993) “Expression, purification, crystallization, and biochemical characterization of recombinant protein phosphatase.” J.Biol.Chem.268:17754-17761 PMID: 8394350