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 mucosa

Reagents 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°C

Storage Conditions

10 mM Tris-HCl
50 mM KCl
1 mM MgCl₂
50% Glycerol
0.1 mM ZnCl₂
pH 8.2 @ 25°C

Heat Inactivation

Molecular Weight

Theoretical: 69 kDa

Specific Activity

3,000 units/mg

Unit 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

温馨提示：不可用于临床治疗。