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| Product Name | Alkaline Phosphatase, Calf Intestine, Recombinant (CIAP, 30 U/uL) |
| Catalog No. | IVDRM-0457 |
| Description | This calf intestine alkaline phosphatase, produced recombinantly in yeast carrying the expression plasmid, removes 5 prime phosphate groups from DNA and RNA and hydrolyses almost all phosphomonoesters without touching phosphodiesters or phosphotriesters. In molecular cloning the enzyme is applied to dephosphorylate digested vectors so that they cannot self ligate, raising the insertion rate of target fragments, and it also prepares DNA templates for 5 prime end labeling. Heat treatment at 65°C for 30 minutes irreversibly destroys 99 percent of activity and phenol abolishes it completely, simplifying downstream ligation. The preparation is free of exonuclease, endonuclease and RNase contamination. |
| Intended Use | Vector dephosphorylation to prevent self ligation, preparation of templates for 5 prime end labeling and protein dephosphorylation |
| Principle / Technology | Hydrolysis of 5 prime terminal phosphate groups from DNA and RNA |
| Sample Type | Digested vector DNA; DNA and RNA substrates for end labeling |
| Reaction Conditions / Protocol | Heat inactivate at 65°C for 30 minutes for 99% irreversible loss of activity |
| Storage Conditions | -20°C |
| Shelf Life | 18 months |
| Package Specifications | 1000 U |
| Quality Control | Unit defined at pH 9.8, 37°C with pNPP substrate; one unit releases 1 umol nitrophenol per minute |
| Key Features | Recombinant yeast production; heat labile simplifying downstream ligation |
| Purity | Free of exonuclease, endonuclease and RNase contamination |
| Concentration | 30 U/uL |
| Shipping Conditions | Dry ice |
| Recommended Buffer System | Stored in 10 mM Tris-HCl pH 8.0, 1 mM MgCl2, 50 mM KCl, 0.1 mM ZnCl2, 50% glycerol |
| Readouts / Downstream Assays | Insertion rate after vector dephosphorylation |
For in vitro diagnostic use.
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