Alkaline Phosphatase Chemical Properties

Density 

1.12 g/mL at 20 °C

storage temp. 

2-8°C

form 

suspension

Specific Gravity

1

color 

white

Water Solubility 

It is soluble in water.

Stability:

Stability Incompatible with strong oxidizing agents

EPA Substance Registry System

Alkaline phosphatase (9001-78-9)

Safety Information

Hazard Codes 

B,Xi

Risk Statements 

36/37/38

Safety Statements 

26-36-24/25-22-23

RIDADR 

UN 0029 1.1B

RIDADR 

UN 0030 1.1B

WGK Germany 

3

F 

3-10

TSCA 

Yes

HazardClass 

1.1B

HS Code 

35079090

MSDS

• Language:English Provider:SigmaAldrich

Alkaline Phosphatase Usage And Synthesis

Chemical Properties

solid

Uses

Hydrolyzes 5′-terminal monophosphate (dephosphorylation) from DNA and RNA. Prevents fragments from self annealing. 5′-nucleic acid targeting for probes.

Uses

Alkaline Phosphatase is commonly used to remove the 5′-terminal phosphate from nucleic acids.

Purification Methods

The E.coli supernatant in sucrose (20%, 33mM) in Tris-HCl pH 8.0 is purified through a DEAE-cellulose column and recrystallised. To the column eluates in 0.125M NaCl is added MgCl2 (to 0.01M) and brought to 50% saturation in (NH4)2SO4 by adding the solid (0.20g/mL). The mixture is centrifuged to remove bubbles and is adjusted to pH 8.0 (with 2N NaOH). Saturated (NH4)2SO4 at pH 8.0 is added dropwise until the solution becomes faintly turbid (~61% saturation). It is set aside at ~25o for 1hour (turbidity will increase). The mixture is placed in an ice bath for several minutes when turbidity disappears and a clear solution is obtained. It is then placed in a large ice bath at 0o (~5L) and allowed to warm slowly to room temperature in a dark room whereby crystals are formed appearing as a silky sheen. The crystals are collected by centrifugation at 25o if necessary. The crystalline solutions are stable at ~25o for many months. They can be stored at 0o, but are unstable when frozen. Cysteine at 10-3M and thioglycolic acid at 10-4M are inhibitory. This is reversed on addition of Zn2+ ions. Many organic phosphates are good substrates for this phosphatase. [Molamy & Horecker Methods Enzymol 9 639 1966, Torriani et al. Methods Enzymol 12b 212 1968, Engstrom Biochim Biophys Acta 92 71 1964.] Alkaline phosphatase from rat osteosarcoma has been purified by Me2CO precipitation and chromatography on DEAE-cellulose, Sephacryl S-200, and hydroxylapatite. [Nair et al. Arch Biochem Biophys 254 18 1987.] Phosphoproteins (various). These are purified by adsorbing onto an iminodiacetic acid substituted agarose column to which are bound ferric ions. This chelate complex acts as a selective immobilised metal affinity adsorbent for phosphoproteins. [Muszyfiska et al. Biochemistry 25 6850 1986.]

Alkaline Phosphatase(9001-78-9)Related Product Information

• Magnesium carbonate hydroxide
• FUCHSIN BASIC
• Cupric acetylacetonate
• Ferric acetylacetonate
• PHENYLSELENOL
• COBALT(II) ACETYLACETONATE
• 2,4-PENTANEDIONE, SILVER DERIVATIVE
• TERT-BUTYL ISOCYANIDE
• Tosylmethyl isocyanide
• N-BUTYLISOCYANIDE
• SALCOMINE
• TRIS(2,2,6,6-TETRAMETHYL-3,5-HEPTANEDIONATO)DYSPROSIUM(III)
• COBALT ETHYLENE DIAMINE CHLORIDE
• Tris(2,4-pentanedionato)chroMiuM(III)
• Aluminum acetylacetonate
• DICHLORO(ETHYLENEDIAMINE)PLATINUM(II)
• 1,1,3,3-TETRAMETHYLBUTYL ISOCYANIDE
• METHYL ISOCYANOACETATE