PHOSPHATASE ALKALINE(AP) EIA GRADE 15 MG

General description

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

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.