Highly active recombinant alkaline phosphatase from the marine bacterium Cobetia marina
![Highly active recombinant alkaline phosphatase from the marine bacterium Cobetia marina Highly active recombinant alkaline phosphatase from the marine bacterium Cobetia marina](/upload/iblock/875/trhknJz1.jpg)
Highly active recombinant alkaline phosphatase from the marine bacterium Cobetia marina
Description. Highly active recombinant alkaline phosphatase from the marine bacterium Cobetia marina (CmAP) is a monomer with a molecular weight of 55 kDa. It is a very effective tool for dephosphorylatiîn of linearized vector prior to ligation. It is also able to dephosphorylate various substrates with a phosphate group available for this enzyme. Monomeric CmAP unlike other commercial alkaline phosphatases is a tool for plasmid-frames. Due to very high specific activity it acts as a powerful tool for immunologic research; for the production of recombinant diagnostic antibodies to cell receptors (proteins benchmark); for dephosphorylating DNA and different proteins. In addition, it can be rapidly inactivated in the reaction by heating for 10 min at 65°C.
Source: The enzyme was fully purified from the recombinant E. coli strain.
Storage conditions: 50 mM Tris-HCl (pH 8.5) buffer solution with 100 mM NaCl, 0,01% NaN3, store at - 20 °C. The enzyme is stable for at least 1 year without losing activity.
Specific activity - 3800 units / mg (Tris-HCl buffer, pH ~ 10)
- 13 000 units / mg (Diethanolamine buffer (DEA), pH 10.3)
Unit Definition: One unit of activity is the amount of enzyme required to form 1 mM p-nitrophenol (p-NP) from 4-nitrophenylphosphate (p-NPP) per minute at 37 °C.
Reaction conditions: 100 mM tris-HCl, 0.2M KCl (pH ~ 10) or 1 M diethanolamine (pH 10.3) at 37 °C. The preparation also exhibits 3-25% activity at pH 6-11 in the acetate, carbonate, and glycine buffers at temperatures from 5 to 50 °C.
Inactivation: Alkaline phosphatase completely denatures after heating for 10 min at 65°C.
Concentration: 0.25 mg/ml (according to Bradford method)
1 unit/µl (Tris-HCl buffer, pH ~ 10)
3 units/µl (Diethanolamine buffer (DEA), pH 10.3)
Dilution buffers: A: 50 mM tris-HCl (pH 8.5)
B: 100 mM tris-HCl (pH ~ 10), 0.2 M KCl
C: 1 M diethanolamine (pH 10.3)