Alkaline Phosphatase
ROCHE/APMB-RO - solution, from bovine (calf) intestine, 2 units/μg protein, optimum pH 7.5-9.5
Synonym: ALP; CAP; CIAP; calf intestinal phosphatase; cip; orthophosphoric monoester phosphohydrolase
Product Type: Chemical
| biological source | bovine (calf) intestine |
| form | solution |
| manufacturer/tradename | Roche |
| optimum pH | 7.5-9.5 |
| packaging | pkg of 1,000 U (10713023001 [1 U/μl]) |
| pkg of 1,000 U (11097075001 [20 U/μl]) | |
| parameter | 37 °C optimum reaction temp. |
| Quality Level | 100  |
| shipped in | wet ice |
| specific activity | 2 units/μg protein |
| storage temp. | 2-8°C |
| 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. |
| 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. |
| Other Notes: | For general laboratory use. |
| 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. |
| 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. |
| RIDADR | NONH for all modes of transport |
| WGK Germany | WGK 1 |
| Flash Point(F) | does not flash |
| Flash Point(C) | does not flash |
| activity | specific activity: 2 units/μg protein |
| Storage Temp. | 2-8°C |
| Enzyme Commission (EC) Number | 3.1.3.1 ( BRENDA  | IUBMB ) |
| UNSPSC | 12352204 |