10567752001

Alkaline Phosphatase

EIA grade, from calf intestine

• Synonym(s): AP, Alkaline Phosphatase
• Enzyme Commission number: 3.1.3.1 (BRENDA, IUBMB)

Properties

• biological source: bovine (calf) intestine
• Quality Level: 100
• form: solution
• specific activity: ~3,000 U/mg enzyme protein (+37°C, 4-nitrophenyl phosphate substrate, diethanolamine as buffer)
• packaging: pkg of 1.5 mL (15 mg)
• manufacturer/tradename: Roche
• optimum pH: 9.8
• pH range: 9.0-11.0
• shipped in: wet ice

Description

General description

Orthophosphoric-monoester phosphohydrolase (alkaline optimum)
Amino groups: 8-16 moles/mole enzyme

Packaging

1.5 mL

Application

Alkaline phosphatase can be coupled to other proteins via its amino groups. The enzyme solution can be used directly for conjugation without prior dialysis.

Quality

Purity: single peak (HPLC)

Sequence

Purification procedure
AP from intestine is a dimeric glycoprotein, it is not known, whether the carbohydrate part is separated during purification procedure. Content of amino groupsIn the literature the carbohydrate content is 2 to 20 percent so one should find approx. 70 amino groups/mol.
In addition one researcher determined the amino groups and found 50 to 60 lysins/mol.-SH groups in AP
No information in literature about -SH groups in AP.

Carbohydrate structure
Results of investigation of the carbohydrate structure of several lots of alkaline phosphatase are available.

NH₂- and carbohydrate estimation
Results of NH₂- and carbohydrate estimation are available.

Unit Definition

Unit Conversion: Approx. 4.35 U (EIA grade AP) [37 °C, 4-NPP as substrate, diethanolamine as buffer, pH 9.8] = 1 U [+25 °C, 4-NPP as substrate, glycine as buffer, pH 10.5]
1.0 U [+37 °C, QC assay conditions] = 0.73 U [+30 °C, QC assay conditions] = 0.48 U [+25 °C, QC assay conditions]

Physical form

Solution, ready-to-use, 10mg/ml

Preparation Note

Storage conditions (working solution): The diluted enzyme (1:10 in 3 M NaCl) will give a stability of 6 months at 4 °C.
1:10 dilution in water will give stability for 1 week at 4°C.
Do not freeze the diluted or undiluted enzyme, because freezing will destroy enzyme activity.

Storage and Stability

Store at 2 to 8 °C. (Do not freeze)

Other Notes

For life science research only. Not for use in diagnostic procedures.

Safety Information

• Storage Class Code: 12 - Non Combustible Liquids
• WGK: WGK 1
• Flash Point(F): does not flash
• Flash Point(C): does not flash