Alkaline Phosphatase, E. coli

Source

Escherichia coli

Applications

• More thermal stable than Calf Intestine Alkaline Phosphatase (CIP)
  
• Optimal incubation temperature is approximately 60°C, however the enzyme remains active from 20°C to 80°C.
  
• Resistant to chemical changes and active over a broad range of buffer conditions.
  
• Can be used to remove 5'-phosphates from DNA or RNA prior to 5'-end labeling (1).
  
• Works to remove 5'-phosphates from linearized vector molecules to prevent self-ligation of the vector during cloning procedures (1).
• Ideal for diagnostic immunoassays and immunodetection of proteins and nucleic acids following blotting experiments (1).

Unit Definition

One unit is the amount of enzyme required to hydrolyze 1 mol of p-nitrophenylphosphate 37°C for one (1) minute in a buffer consisting of:

0.25 mM MgCl₂ (pH 9.8).
1 M diethanolamine.
10 mM p-nitrophenylphosphate.

Storage Buffer

20 mM Tris-HCl (pH 7.0 at 22°C)
5 mM potassium phosphate
100 mM KCl
0.1 mM MgCl₂
0.1 mM ZnCl₂  
+Stabilizers

Quality Control

All preparations are assayed for contaminating endonuclease and nonspecific RNase and single- and double-stranded DNase activities

Storage Conditions

Store at -20°C
Product shipped on dry ice