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| 货号： | M0290 |
| 保存条件： | -20 |
| 规格： | V S L |
Alkaline Phosphatase, Calf Intestinal (CIP)
| M0290L | Alkaline Phosphatase, Calf Intestinal 小牛肠碱性磷酸酶 (CIP) | 5,000 units |
| M0290S | Alkaline Phosphatase, Calf Intestinal 小牛肠碱性磷酸酶 (CIP) | 1,000 units |
| M0290S | Alkaline Phosphatase, Calf Intestinal 小牛肠碱性磷酸酶 (CIP) | 500 units |
Description
Alkaline Phosphatase, Calf Intestinal (CIP) nonspecifically catalyzes the dephosphorylation of 5´ and 3´ ends of DNA and RNA phosphomonoesters. Also, CIP hydrolyses ribo-, as well as deoxyribonucleoside triphosphates (NTPs and dNTPs). CIP is useful in many molecular biology applications such as the removal of phosphorylated ends of DNA and RNA for subsequent use in cloning or end-labeling of probes. In cloning, dephosphorylation prevents religation of linearized plasmid DNA. The enzyme acts on 5´ protruding, 5´ recessed and blunt ends. CIP may also be used to degrade unincorporated dNTPs in PCR reactions to prepare templates for DNA sequencing or SNP analysis.Product Source
Calf intestinal mucosaReagents Supplied
The following reagents are supplied with this product:
| Store at (°C) | Concentration | |
| CutSmart™ Buffer | -20 | 10X |
Advantages and Features
Applications
- Dephosphorylation of cloning vector DNA to prevent recircularization during ligation
- Dephosphorylation of DNA prior to end-labeling using T4 Polynucleotide Kinase
- Treatment of dNTPs in PCR reactions prior to sequencing or SNP analysis
- Dephosphorylation of DNA and RNA
Properties and Usage
Unit Definition
One unit is defined as the amount of enzyme that hydrolyzes 1 μmol of p-Nitrophenyl Phosphate, PNPP (NEB #P0757) in a total reaction volume of 1 ml in 1 minute at 37°C.Reaction Conditions
1X CutSmart™ Buffer
Incubate at 37°C
1X CutSmart™ Buffer:
50 mM Potassium Acetate
20 mM Tris-acetate
10 mM Magnesium Acetate
100 μg/ml BSA
pH 7.9 @ 25°C
Storage Temperature
-20°CStorage Conditions
10 mM Tris-HCl
50 mM KCl
1 mM MgCl2
50% Glycerol
0.1 mM ZnCl2
pH 8.2 @ 25°C
Heat Inactivation
NoMolecular Weight
Theoretical: 69 kDaSpecific Activity
3,000 units/mgUnit Assay Conditions
1 M diethanolamine- HCl (pH 9.8), 0.5 mM MgCl2, 50 mM p-nitrophenylphosphate and enzyme. These conditions are only used for quantitating enzyme activity.Notes
- CIP, as are most alkaline phosphatases, is a Zn2+ and Mg2+-dependent enzyme. Our formulation of its storage buffer provides Zn2+ and Mg2+, which does not require supplemental zinc or other additives in reactions with CIP.
- CIP is also active in 1X NEBuffers 1.1, 2.1, 3.1, as well as NEBuffers 1, 2, 3, 4 and unique NEBuffer for EcoRI.
- CIP activity is enhanced in the presence of monovalent salts.
- CIP is inhibited by metal chelators (e.g. EDTA), inorganic phosphate and phosphate analogs.
References
- Sambrook, J., Fritsch, E.F. and Maniatis, T. (1989). Molecular Cloning: A Laboratory Manual, (2nd ed,). (p5.72), Cold Spring Harbor Laboratory Press .
- Mossner, E., Boll, M. and Pfleiderer, G. (1980). Hoppe Seylers Z. Physiol. Chem.. 361
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