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APMB-RO

Roche

Alkaline Phosphatase

solution, from bovine (calf) intestine, 2 units/μg protein, optimum pH 7.5-9.5

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Synonym(s):
ALP, CAP, CIAP, calf intestinal phosphatase, cip, orthophosphoric monoester phosphohydrolase
Enzyme Commission number:

biological source

bovine (calf) intestine

Quality Level

form

solution

specific activity

2 units/μg protein

packaging

pkg of 1,000 U (10713023001 [1 U/μl])
pkg of 1,000 U (11097075001 [20 U/μl])

manufacturer/tradename

Roche

parameter

37 °C optimum reaction temp.

optimum pH

7.5-9.5

shipped in

wet ice

storage temp.

2-8°C

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This Item
10567752001RAP-RO539514
Alkaline Phosphatase solution, from bovine (calf) intestine, 2 units/μg protein, optimum pH 7.5-9.5

Roche

APMB-RO

Alkaline Phosphatase

Alkaline Phosphatase EIA grade, from calf intestine

Roche

10567752001

Alkaline Phosphatase

rAPid Alkaline Phosphatase 1 units/μg protein, optimum reaction temp. 37 °C

Roche

RAP-RO

rAPid Alkaline Phosphatase

form

solution

form

solution

form

-

form

liquid

specific activity

2 units/μg protein

specific activity

~3,000 U/mg enzyme protein (+37°C, 4-nitrophenyl phosphate substrate, diethanolamine as buffer)

specific activity

1 units/μg protein

specific activity

≥400,000 units/mL

manufacturer/tradename

Roche

manufacturer/tradename

Roche

manufacturer/tradename

Roche

manufacturer/tradename

Calbiochem®

optimum pH

7.5-9.5

optimum pH

9.8

optimum pH

-

optimum pH

-

shipped in

wet ice

shipped in

wet ice

shipped in

-

shipped in

wet ice

General description

Alkaline phosphatase (ALP) is a glycoprotein that is expressed ubiquitously. This enzyme is bound to the cell membrane. Activation of ALP requires essential co-factors, such as zinc and magnesium. Alkaline phosphatase catalyzes the removal of phosphate groups from various compounds that are phosphorylated. Alkaline phosphatase from calf intestine hydrolyzes 5′-monophosphate groups from both DNA and RNA. It can also hydrolyze 5′-diphosphate and 5′-triphosphate groups from RNA. Hence, intestinal ALP is one of the commonly used enzymes in molecular cloning.

Application

Use this preparation of calf intestinal alkaline phosphatase to remove 5′-terminal phosphates from DNA or RNA and immunoprecipitated RNA samples.
Note: The 11097075001 preparation has a high concentration of enzyme (20 U/μl), which is convenient for large-scale experiments. The 10713023001 preparation is a lower concentration (1 U/μl), which is convenient for small-scale experiments.

Sequence

AP is a zinc-containing enzyme with 4 atoms zinc per molecule (Efstradiatis, 1977).

Unit Definition

One unit of alkaline phosphatase is the enzyme activity which hydrolyzes 1 mol of 4-nitrophenyl phosphate in 1 minute at +37 °C under assay conditions.
Note: According to Moessner et al. 5 units alkaline phosphatase (+37 °C; diethanolamine buffer) correspond to 1 unit alkaline phosphatase (+25 °C; glycine/NaOH buffer).

Unit Conversion: Approx. 3.6 U (MB 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].

Volume Activity: 20 U/μl for the 11097075001 preparation. 1 U/μl for the 10713023001 preparation.

Physical form

10713023001: Enzyme solution (1 U/μl) in storage buffer, pH 7.6 (20 °C)

11097075001: Enzyme solution (20 U/μl) in storage buffer, pH 7.6 (20 °C)

Preparation Note

Working solution: Storage buffer: 25 mM Tris-HCl, 1 mM MgCl2, 0.1 mM ZnCl2, glycerol 50% (v/v), pH 7.6 (4 °C).
Storage conditions (working solution): Dilutions (1 to 5 U/μl) of MB grade AP in the storage buffer are stable for 17 months at 2 to 8 °C.
Higher Dilutions (~0.01 U/μl) should be made fresh daily and kept at 2 to 8 °C.
Preventing self-ligation of vectors during cloning
If a linearized vector lacks a 5′ phosphate, it cannot ligate to itself or form concatamers that contain multiple copies of the vector. Thus, the product of the ligation reaction is predominantly recombinant DNA (vector + DNA insert), rather than religated plasmid.
Inactivation: Add 1/10 volume of EGTA to the reaction mix and incubate at +65°C for 10 minutes. To ensure complete inactivation of enzyme, extract the mix with phenol/chloroform/isoamyl alcohol to remove all protein.

Other Notes

For general laboratory use.

Storage Class Code

12 - Non Combustible Liquids

WGK

WGK 1

Flash Point(F)

does not flash

Flash Point(C)

does not flash


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Alkaline Phosphatase
Lowe D, et al
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Alkaline Phosphatase.
Green, Michael R and Sambrook, Joseph
Cold Spring Harbor Protocols, 100768-100768 (2020)
Qiang Zhao et al.
Physiologia plantarum, 174(4), e13731-e13731 (2022-06-20)
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O Brenna et al.
The Biochemical journal, 151(2), 291-296 (1975-11-01)
A crude preparation of alkaline phosphatase (EC 3.1.3.1) from calf intestinal mucosa was purified by affinity chromatography on Sepharose-bound derivatives of arsanilic acid, which was found to be a competitive inhibitor of the enzyme. Three biospecific adsorbents were prepared for

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