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Alkaline Phosphatase Calf Intestinal(CIAP) / 1KU / Cat.#3002
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  Alkaline Phosphatase,

  Calf Intestinal (CIAP)

    

#3002

 

1,000 units

#3002L

 

5,000 units

 

 

DESCRIPTION 

Alkaline Phosphatase catalyszes the removal of 5'-phosphate groups from DNA, RNA and ribo- and deoxyribonucleoside triphosphates.  

 

SOURCE 

Calf intestinal mucosa. 

 

APPLICATIONS 

¡Ü Removing 5' phosphates from DNA or RNA 

prior to end labeling 5¡¯ termini with 32P

¡Ü Removing 5' phosphates from fragments of 

   DNA to prevent self-ligation. 

 

UNIT DEFINITION 

One unit is defined as the amount of enzyme required to catalyze the hydrolysis of 1 ¥ìmol of 4-nitrophenyl- phosphate to 4-nitrophenol in 1 minute at 37¡É. 


NOTE
 

Proteinase K is used to digest CIAP, which must be completely removed if subsequent ligatios are to work efficiently. An alternative method is to activate the CIAP by heating to 65¡É for 1 hour(or 75¡É for 10 minutes) in the presence of 5 mM EDTA(pH 8.0) and then to purify the dephosphorylated DNA by extraction with phenol:chloroform.


 

 

 

STANDARD UNIT ASSAY CONDITIONS 

1 M diethanolamine (pH 9.8), 0.5 mM MgCl2,  10.9 mM 4-nitrophenylphosphate. 

 

CONCENTRATION: 1-5 unit/§¡

 

10X REACTION BUFFER 

500 mM Tris-HCl (pH 9.3), 10 mM MgCl2, 1 mM ZnCl2, 10 mM spermidine. 

 

STORAGE CONDITIONS 

10 mM Tris-HCl (pH £Ü8.0), 1 mM MgCl2, 0.1 mM ZnCl2, 50 mM KCl and 50% glycerol.  Store at -20¡É.¡¡  

 

REFERENCE 

1. Sambrook, J. and Russell, David W. (2001) Molecular Cloning: A Laboratory Manual, 3rd ed., Cold Spring Harbor Laboratory Press. 

 


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