Shrimp Alkaline Phosphatase
ArcticZymes’ Shrimp Alkaline Phosphatase (SAP) is the only heat labile, all-purpose alkaline phosphatase purified from a recombinant source and originally isolated from Pandalus borealis (arctic shrimp). The recombinant production leads to increased storage stability, low batch-to-batch variations and high specific activity. For added flexibility, when lyophilization may be desired, SAP is also available in a glycerol-free format.
SAP is useful in many molecular biology applications by offering fast and easy dephosphorylation of DNA, RNA and nucleotides. SAP is also active in most restriction enzyme buffers and is completely and irreversibly inactivated after 5 minutes at 65°C (figure 1). This property simplifies most workflows involving alkaline phosphatase treatment.
It has been on the market since 1993 and is now replaced by the new and better recombinant form. The Recombinant enzyme is produced in Pichia Pastoris, and has all the properties of the well-proven native version, but with additional benefits. The recombinant version is far more stable at ambient temperature, is also of high, consistent purity, and available at larger batches at high concentration
100% heat-inactivated at 65°C
Works in restriction enzyme buffers
Used in PCR clean-up
Removes 5’-phosphates from DNA, RNA, dNTPs and dephosphorylates proteins
Also available in Glycerol-free formulation
Unit definition: One Unit will convert 1 µmol of p-nitrophenyl phosphate per minute to nitrophenol and phosphate at 37ºC and pH 10.4 in 0.1 M glycine buffer, 1 mM each of ZnCl2 and MgCl2 and 6 mM 4-nitrophenyl phosphate. This Unit definition makes 1 Unit of SAP equivalent to 5 to 40 Units of Antarctic Phosphatase (New England Biolabs), 1.3 Units APex phosphatase or 35 Units NTPhos phosphatase (Epicentre Biotechnologies). SAP is therefore the most processive alternative.
- Specific activity: >2 000 Units/mg.
- Purity: DNase activities not detected.
- Concentration: Minimum 10 000 Units/ml, available up to 30 000 Units/ml.
- Batch sizes: 25 – 35 million Units / batch.
Stable at -20ºC in storage buffer (25 mM Tris-HCl pH 7.6, 5 mM MgCl2, glycerol 50%). At room temperature, >95% activity remains after 90 days.
Still, the enzyme is completely inactivated after 5 minutes at 65°C. At 75°C, SAP is completely inactivated after only 1 minute. In a standard thermocycler, the process of heating from 37 to 95 and back to 37°C is sufficient to completely inactivate SAP.
Recombinant SAP is covered by the following patents: US 7,323,325, EP 1,326,890, JP 4,191,479, AU 9,398,701 and related applications in other countries either granted or pending.
Dephosphorylation of cloning vectors
- Generating In Vivo Cloning Vectors for Parallel Cloning of Large Gene Clusters by Homologous Recombination. Lee J, Rha E, Yeom SJ, Lee DH, Choi ES and Lee SG. PLoS One. 2013; 8(11): e79979.
Degradation of nucleotides before genotyping or sequencing
- Comprehensive evaluation of coding region point mutations in microsatellite‐unstable colorectal cancer. Kondelin J, Salokas K, Saarinen L, Ovaska K, Rauanheimo H, Plaketti RM, Hamberg J, Liu X, Yadav L, Gylfe AE, Cajuso T, Hänninen UA, Palin K, Ristolainen H, Katainen R, Kaasinen E, Tanskanen T, Aavikko M, Taipale M, Taipale J, Renkonen-Sinisalo L, Lepistö A, Koskensalo S, Böhm J, Mecklin J-P, Ongen H, Dermitzakis ET, Kilpivaara O, Vahteristo P, Turunen M, Hautaniemi S, Tuupanen S, Karhu A, Välimäki N, Varjosalo M, Pitkänen E, Aaltonen LA.
EMBO Mol Med. 2018; 10(9): e8552.
- Molecular systematics of the subfamily Limenitidinae (Lepidoptera: Nymphalidae).
Dhungel B., Wahlberg N. PeerJ. 2018; 6, e4311.
- Detection and characterization of a rhabdovirus causing mortality in black bullhead catfish, Ameiurus melas. Bedendo G, Panzarin V, Fortin A, Zamperin G, Pretto T, Buratin A, Quartesan R, Sabbion M, Salogni C, Pascoli F, Toffan A. J Fish Dis. 2018; 41 (7):1063-1075.
- Pellets of proof: First glimpse of the dietary composition of adult odonates as revealed by metabarcoding of feces. Kari M. Kaunisto, Tomas Roslin, Ilari E. Sääksjärvi, Eero J. Vesterinen. Ecol Evol. 2017; 7:8588–8598.
- Systematics and origin of moths in the subfamily Arctiinae (Lepidoptera, Erebidae) in the Neotropical region. Zenker MM, Wahlberg N, Brehm G, Teston,JA, Przybylowicz L, Pie MR, Freitas AVL. Zoologica Scripta. 2016; 46 (3): 348-362.
- Medium-throughput SNP genotyping using mass spectrometry: multiplex SNP genotyping using the iPLEX® Gold assay. Millis MP. Methods Mol Biol. 2011; 700:61-76.
- Dried reagents for multiplex genotyping by tag-array minisequencing to be used in microfluidic devices. Ahlford A, Kjeldsen B, Reimers J, Lundmark A, Romani M, Wolff A, Syvänen AC, Brivio M. Analyst. 2010; 135(9): 2377-85.
- Highly multiplexed genotyping of thiopurine s-methyltransferase variants using MALD-TOF mass spectrometry: reliable genotyping in different ethnic groups. Schaeffeler E, Zanger UM, Eichelbaum M, Asante-Poku S, Shin JG, Schwab M. Clin Chem. 2008; 54(10): 1637-47.
- Genotyping SNPs using a UV-photocleavable oligonucleotide in MALDI-TOF MS. Vallone PM, Fahr K, Kostrzewa M. Methods Mol Biol. 2005; 297: 169-78.
- A gel-free SNP genotyping method: bioluminometric assay coupled with modified primer extension reactions (BAMPER) directly from double-stranded PCR products. Zhou GH, Shirakura H, Kamahori M, Okano K, Nagai K, Kambara H. Hum Mutat. 2004; 24(2): 155-63.
- G2 checkpoint in uterine cervical cancer with HPV 16 E6 according to p53 polymorphism and its screening value. Cho NH, Lim SY, Kim YT, Kim D, Kim YS, Kim JW. Gynecol Oncol. 2003; 90(1): 15-22.
- A novel procedure for efficient genotyping of single nucleotide polymorphisms. Sauer S, Lechner D, Berlin K, Lehrach H, Escary JL, Fox N, Gut IG. Nucleic Acids Res. 2000; 28(5): E13.
- Autophosphorylation activates Dictyostelium myosin II heavy chain kinase A by providing a ligand for an allosteric binding site in the alpha-kinase domain.Crawley SW, Gharaei MS, Ye Q, Yang Y, Raveh B, London N, Schueler-Furman O, Jia Z, Côté GP. J Biol Chem. 2011; 286(4): 2607-16.
- A diurnally regulated dehydrin from Avicennia marina that shows nucleo-cytoplasmic localization and is phosphorylated by Casein kinase II in vitro. Mehta PA, Rebala KC, Venkataraman G, Parida A. Plant Physiol Biochem. 2009; 47(8): 701-9.
SAP for quantification
- Dephospho-CoA kinase provides a rapid and sensitive radiochemical assay for coenzyme A and its thioesters. Wadler C, Cronan JE. Anal Biochem. 2007; 368(1): 17-23.
- A comparison of enzymatic digestion for the quantitation of an oligonucleotide by liquid chromatography-isotope dilution mass spectrometry. Donald CE, Stokes P, O’Connor G, Woolford AJ. J Chromatogr B Analyt Technol Biomed Life Sci. 2005; 817(2): 173-82.
SAP for pyrosequensing
- Method enabling pyrosequencing on double-stranded DNA. Nordström T., Nourizad K., Ronaghi M., Nyrén P. Anal Biochem. 2000; 282(2): 186-93.
- Sensitive electrochemical assay of alkaline phosphatase activity based on TdT-mediated hemin/G-quadruplex DNAzyme nanowires for signal amplification. (ScienceDirect pay service). Liu Y, Xiong E, Li X, Li J, Zhang X, Chen J Biosensors and Bioelectronics. 2017; 87: 970-975.
- Ligand-binding and metal-exchange crystallographic studies on shrimp alkaline phosphatase. de Backer MM, McSweeney S, Lindley PF, Hough E. Acta Crystallogr D Biol Crystallogr. 2004; 60(Pt 9):1555-61.
- Thermolabile alkaline phosphatase from Northern shrimp (Pandalus borealis): protein and cDNA sequence analyses. Nilsen IW, Øverbø K, Olsen RL. Comp Biochem Physiol B Biochem Mol Biol. 2001; 129(4): 853-61.